PRE-MEDICAL CAREER POINT

CA REER POINT

PRE- MEDICAL DIVISION

GENETICS

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GENETICS
Genetics term was given by W.Bateson. (1905) (Father of Modern Genetics).
Genetics = Branch of biology which deals with the study of heredity and variation
 Heredity – Transmission of genetic characters from generation to generation.
 Variation – Individuals of same species have some difference, these are called variation.

Variation

Somatic variation Germinal (Blastogenic) variation

(Non inheritable) transfer from generation to generation (Inheritable)
Two types

Continuous variation Discontinuous variation

(due to crossing over) (due to mutation)

Substantive Meristic Substantive Meristic

 INHERITANCE : HEREDITY AND VARIATIONS
Heredity : It is the transmission of genetic characters from parents to the offsprings. It deals with the
phenomenon of ‘‘like begets like’’ e.g., human babies are like human beings in overall characteristics.
Variations are common in sexually reproducing organisms. Variations are of following two types :
 Somatogenic : These are acquired variations and are non-inheritable in nature. The ability of an
organism to alter its phenotype in response to environment is called phenotypic plasticity.
 Blastogenic Variations : These are germinal variations and are hereditary in nature. They are again
of two types:
 Continuous variations : These are the fluctuating variations and can not give rise to new species.
These are further of two types
 Substantive : Variation in size, shape and colour of organism.
 Meristic : Variation in number of parts e.g. number of grains in an ear of wheat
 Discontinuous Variations : Also known as mutations, sports or saltations. These variation are
responsible for formation of new species and organism thus formed is called mutant.
Types of discontinuous variations
 Substantive variations : These influence shape, colour, size etc., e.g., hairless cat, short
legged ancon sheep.
 Meristic variation : These affect number of parts e.g. polydactyly in humans.
Variations are significance in evolution as they make the organism better suited to modifying
environmental conditions, produce new trait in organism and provide raw material for evolution.

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History :
 Muller : Proposed the term ‘‘Cytogenetics’’ (Cytology + Genetics)
Father fo Actinobiology
Actinobiology - study the effect of radiation of living organisms.
 Morgan : Father of Experimental genetics
He experiment on Drosophila melanogaster & proposed various concepts
Gene theory : According to gene theory; genes are linearly located on chromosome.
Linkage term, Theory of sex linkage, Crossing over term, Criss - cross inheritance.

EARLY SPECULATIONS (PREMENDELIAN)

(1) Vapour Theory (2) Preformation Theory
(3) Encasement Theory (4) Theory of Epigenesis
(5) Pangenesis Theory (6) Weismann Theory of Germ plasm
(7) Mendelism
(1) Vapour Theory :
 Proposed by a Greek philosopher Pythagoras in 500 B.C.
 Each organ of an animal body emitted some kind of vapour and that a new individual was formed by
combination of the vapour from different organs.
(2) Preformation Theory (1632 – 1723) :
 Dutch scientist ‘Swammerdam’ propounded the preformation theory. This theory holds that the sex
cells (sperm and ova) had the miniature copy of adults and the development of embryo was actually
only the enlargement of parts that were already present in the sperm or egg.
 This minature form of the animal present in the gamete was called ‘Homunculus’.
 This theory was supported by Hartsoeker.
 Those who considered homunculus to be present in sperms were called Spermist.
 Those who considered homunculus to be present in ova (eggs) were called Ovist.
 Antony Van Leeuwenhoek was the first to observe human sperms.
(3) Encasement Theory :
‘Charles Bonnet’ and his supporters presumed that every female contains within her body minature
prototypes of all the creatures which will every descend from her, one generation within the other some
what like a series of chinese bones. (a box inside a box and so on).
(4) Theory of Epigenesis :
 Wolff proposed that germ cells contain definite but undifferentiated substances which, after fertilization
becomes organized into various complex body organs (Differentiated) which form the adult.
This idea was referred to as epigensis.
(5) Pangenesis Theory :
 Charles Darwin propounded pangenesis theory.
 According to this theory every cell, tissue and organ of animal body produces many minute particles
known as pangenes or gemmules.
 These gemmules are discharged in the blood stream and are deposited in the reproductive organs.
These reproductive cells contain these pangenes and a child develops as a result of blending of the
pangenes from two parents.
 Thus the individual would represent the mixture of both of the parents.
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(6) Theory of Germplasm :
 ‘August Weismann’ (1889) suggested the theory of continuity of germplasm.
 They referred to the reproductive cells as germplasm and to rest of the body as somatoplasm.
 The germplasm forms the bridge of life and passes from one generation to the next.
 Soma is ‘Mortal’ because it eventually dies and disintegrates, the germ is “immortal”, because it
continues forever.
MENDELISM :
Experiments performed by Mendel on genetics and description of mechanisms of hereditery processes
and formulation of principles are known as Mendelism.
Gregor Johann Mendel (1822 – 1884) : Mendel was born on july 22, 1822 at Heinzendorf in Austria
a Silesia village. Mendel worked in Augustinian Monastery as monk at Brunn city, Austria.
In 1856-57, he started his historical experiments of heredity on pea (pisum sativum) plant. His
experimental work continued on pea plant till 1865 (19th century)
The results of his experiments were published in the science journal, “Nature For schender varien”
in 1866.
This journal was in Germen language. Title was “verschue uber Pflangen Hybridan”.
This journal was published by ‘Natural History society of Bruno’.
A paper of Mendel by the name of ‘‘Experiment in plant Hybridization’’ published in this journal. Mendel
was unable to get any popularity. No one understood of him. He died in 1884 without getting any credit
of his work due to kidney disease (Bright disease)
After 16 years of Mendel’s death in 1900, Mendel’s postulates were rediscovered. Rediscovery by three
scientists independently.
 Carl Correns : Germany (Experiment on Maize)
 Hugo deVries (Holland) (Experiment on Evening Primerose)
He republished the Mendel’s results in 1901 in Flora magazine
 Erich von Tschermak Seysenegg - (Austria) (Experiment on different flowering plants)
The credit of rediscovery of Mendelism goes to three scientists.
Correns given two laws fo Mendelism
Law of heredity / Inheritance / Mendel
 Mendel Results Remain Hidden due to :
 At that time Darwin’s book ‘‘Origin of Species’’ published. Scientists were busy in discussion with this
book.
 Medel’s ideas were ahead of that time.
 Mendel used higher statistical calculation in his experiments so the results were complicated to
understand.
 Mendel also performed his experiments on Hawkeweed (hieracium) and beans (Lablab) plant on suggestion
of Karl Nageli but mendel did not get succeed in Heriacium, Parthenogenesis is present in it.
 Reasons for Mendel’s success :
 Mendel studied the inheretance of one or two characters at a time unlike his predecessors who had
considered many characters at a time (Kolreuter-Tobacco plant, John Goss & Knight-Pea plant).

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 Selection of Material (Pea plant - Pisum sativum)
 Selection of garden pea plant is suitable for studies. Which have following advantage.
 Pea plant is annual plant with short life cycle of 2-3 months so large number of offsprings can be
analysed within a short period of time.
 It has many contrasting traits.
 Natural self pollination is present in pea plant so purity of character is maintained in it.
 Cross pollination can be performed in it artificially so hybridization can be made possible.
 Pea plant easy to cultivate.
 Pea seeds are large. In addition to pea, Mendel worked on rajama.
 Mendel quantitatively analyse the inheritance of qualitative characters.
 He maintained the statistical records of all the experiments.
 Mendel’s work : Mendel studied 7 characters or 7 pairs of contrasting traits.
Actual data obtained by Mendel in F2 progenies in garden pea
Character (Chromosomal
S.No. Dominant Recessive Ratio
position)
1 Length of plant (4) 787 (tall) 277 (dwarf) 2.84:1
2 Colour of flower (1) 705 (violet) 224 (white) 3.15:1
3 Pod or flower position (4) 651 (axial) 207 (terminal) 3.14:1
4 Shape of pod (4) 882 (inflated) 299 (constricted) 2.94:1
5 Colour of pod (5) 428 (green) 152 (yellow) 2.82:1
6 Shape of seed (7) 5,474 (round) 1850 (wrinkled) 2.96:1
7 Colour of cotyledon (1) 6,022 (yellow) 2,001 (green) 3.01:1
Average of all traits studied 2.98:(=3:1)

 Technique of Mendel
He developed a technique Emasculation and Bagging for hybridization in plants.
Flowers of pea plant are bisexual. In this method one considered as male and another as female.
The plant used as female, stamens of this plant are removed at juvenile stage, this is called Emasculation.
Emasculation is done to prevent self pollination.
Emasculated flowers covered by bags, this is called bagging.
Bagging is only used to prevent undersirable cross pollination.
Mature pollen grains are collected from male plants and spread over emasculated flower.
Seeds are formed in the female flower after pollination.
The plants that are obtained from these seeds are called First Filial generation or F1 generation according
to mendel.
Mendel was great plant breader (true breader)
 SOME DEFINED TERMS :
 Factors : Unit of heredity which is responsible for inheritance and appearance of characters. These
factors were referred as genes by Johannsen (1909). Mendel used term “element” for factor.

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 Morgan first use symbol to represent the factor. Dominant factor are represented by capital letter while
recessive factor by small letter
 Allele : Alternative forms of a gene which are located on same position [locus] on the homologous
chromosome is called Allele. Term allele was coined by Bateson.

T T T t t t

 Homozygous : A zygote is formed by fusion of two gametes having identicle factors is called homozygote
and organism developed from this zygote is called homozygous. Ex. TT, RR, tt
 Heterozygous : A zygote is formed by fusion of two different types of gamete carrying different factors
is called Heterozygous (Tt) (Rr) and individual developed from such zygote is called hetrozygous. The
term homozygous and heterozygous are coined by Bateson.
 Hemizygous : If individual contains only one gene of a pair then individual said to be hemizygous. Male
individual is always hemizygous for sex linked gene.
 Phenotype : It is the external and morphological appearance of an organism for a particular character.
 Genotype : The genetic constitution or genetic make-up of an organism for a particular character.
Genotype & phenotype terms were coined by Johannsen.
 Phenocopy : If different genotypes are placed in different environmental conditions if they produce
same phenotype. These individual are said to be Phenocopy of each other.

MONOHYBRID CROSS

When we consider the inheritance of one character at a time by a cross is called monohybrid cross.
First of all, Mendel selected tall and dwarf plants

Tall Dwarf
(Pure) (Pure)

F1-Generation
All tall (impure)
Self pollination (Selfing)

F2-Generation Tall Dwarf(Phenotypic ratio or basic ratio or Mendelian ratio)

3 1

Self Pollination
1 Tall (Pure) 2 Tall (impure)
(Selfing) (Selfing)
Dwarf (pure)
F3-Generation All Tall 3 Tall : 1 dwarf

Conclusions (results) of Monohybrid Cross

st
I Conclusion :
According to Mendel each genetic character is controlled by a pair of unit factor. It is known as
conclusion of paired factor or unit factor.
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nd
II Conclusion :
This conclusion is based on F1 - generation. When two different unit factors are present in single
individual, then only one unit factor is able to express itself and known as dominant unit factor. Another
unit factor fails to express is the recessive factor. In the presence of dominant unit factor recessive unit
factor can not express and it is known as conclusion of dominance.
Tall Dwarf
TT tt

F1-Generation Tt All tall

rd
III Conclusion :
During gamete formation; the unit factors of a pair segregate randomly and transfer inside different gamete.
Each gamete receives only one factor of a pair; so gametes are pure for a particular trait. It is known as
conclusion of purity of gametes or segregation.
Tt
gametogenesis

Tt Tt
gamete gamete
DIHYBRID CROSS
A cross in which study of inheritance of two pairs of contrasting traits or two characters.
Mendel wanted to observe the effect of one character on another character.
Mendel selected traits for dihybrid cross for his experiment as follows :
 Colour of cotyledons  Yellow (Y) & Green (y)
 Seed form  Round (R) and Wrinkled (r)
yellow and round characters are dominant and green and wrinkled are recesive characters. Mendel crossed
yellow and round seeded plants with green and wrinkled seeded plants. All the plants in F1-generation had
yellow and round seeds.
All the plants in F1-generation had yellow and round seeds.
When F1 plants were self pollinated to produce four kinds of plants in F2 generation such as yellow round,
yellow-wrinkled, green round and green wrinkled, there were in the ratio of 9 : 3 : 3 : 1. This ratio is known
as Dihybrid ratio.
Yellow Round Green Wrinkled

F1-Generation

All yellow round

selfing

Yellow Round Green Round Yellow Wrinkled Green Wrinkled

9 : 3 : 3 : 1
New Combination
Expression of yellow round (9) and green wrinkled (1) traits shows as their parental combination. Green
round and yellow wrinkled type of plants are produced by the results of new combination (Recombinant).
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 Conclusion :
The F2 generation plant produce two new phenotypes, so inheritance of seed coat colour is independent
from the inheritance of shape of seed. Otherwise It can not possible to obtain yellow wrinkled and green
round type of seeds.
This observation leads to the Mendel’s conclusion that different type of characters present in plants
assorted independently during inheritace.
This is known as Conclusion of Independent Assortment. It is based on F 2 - generation of dihybrid
cross. The nonhomologous chromosome show random distribution during anaphasei-I of meiosis.
 Explaination :
A pure yellow and round seeded plant crossed with green and wrinkled seeded plant which are having
genotype YYRR and yyrr to produced F1 generation having YyRr genotype.
Both the characters recombine independently from each other during gamete formation in F 1 generation.
Factor (R) of pair factor (r) is having equal change to (Y) factor or (y) factor of gametes during recombination
to form two type of gametes (YR) and (yr).
Similarly (r) factor also having equal change with (Y) factor or (y) factor of gametes to form a two type
gamets (Yr) and (yr).
Thus, total four types of gametes - (YR), (yr), (yR), (yr) are formed.
Therefore during the gametes formation in F1 generation new combination or recombination appear in F2.
 Demonstration by checker board method
Yellow Round Yellow Round

Parents  YY RR yy rr

Gametes  YR yr

F1-Generation  Yy Rr

Self Pollination

 F2 - Generation 

YR Yr yR yr

YR YYRR YYRr YyRR YyRr

Yr YYRr YYrr YyRr Yyrr

yR YyRR YyRr yyRR yyRr

yr YyRr Yyrr yyRr yyrr

Yellow Round = 9 / 16
Yellow Wrinkled = 3 / 16
Green Round = 3 / 16
Green Wrinkled = 1 / 16
Phenotypic Ratio = 9 : 3 : 3 : 1
Phenotype Genotype
Homozygous yellow & Homozygous Round — YY RR = 1
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Homozygous yellow & Heterozygous Round — YY Rr = 2
Heterozygous yellow & Homozygous Round — Yy RR = 2
Heterozygous yellow & Heterozygous Round — Yy Rr = 4
Homozygous yellow & Homozygous wrinkled — YY rr = 1
Heterozygous yellow & Homozygous wrinkled — Yy rr = 2
Homozygous green & Homozygous Round — yy RR = 1
Homozygous green & Heterozygous Round — yy Rr = 2
Homozygous green & Homozygous wrinkled — yy rr = 1
F2 Genotypic Ratio = 1:2:2:4:1:2:1:2:1 —

Num be r Num be r of
Type s of Num be r of Num be r of
of tra its Zygote s / P he notypic Ge notypic
Ex pe rim e nt ga m e te s P he notype ge notype
hybrid offsprings Ra tio Ra tio
(2 n ) 2 (2 n ) (3 n )
(n) (ga m e te s)
M onohybrid
1 2n = 21 = 2 22 = 4 2n = 21 = 2 31 = 3 3:1 1: 2: 1
cross (Aa × A a)

(1 : 2 : 1)2 =
Dihybrid c ros s 2 2 2 2 (3 : 1)2 =
2 2 = 4 4 = 16 2 = 4 3 = 9 2:4: 2: 1:2
(AaBb × A aBb) 9 :3: 3: 1
: 1: 1:2:1

Trihybrid cross
AaB bCc
3 23 = 8 8 2 = 64 23 = 8 3 3 = 27 (3 : 1)n (1 : 2 : 1)3
×
AaB bCc

 Back Cross :
A back cross is a cross in which F1 individuals are crossed with any of their parents.
When F1 individual is crossed with dominant parent then it is termed out cross. The generation obtained
from this cross, all possess dominant character.
 Test Cross : When F 1 progency is crossed with recessive parent then it is called test cross. The total
generations obtained from this cross, 50% having dominant character and 50% having recessive character.
[Monoybrid test cross]. Test cross helps to find out the genotype of dominant individual. Whether it is
homozyous or heterozyous for that character
 Monhybrid Test Cross : The progeny obtained, from the monohybrid test cross are in equal proportion,
means 50% is dominant phenotypes and 50% is recessive phenotypes. It can be represented in
symbolic forms as follows.
F1 progeny(hybrid) F recessive parent
Tt × tt

t
T Tt
Monohybrid test cross ratio = 1 : 1
t tt

 Dilhybrid Test Cross : The progency is obtained from dihybrid test cross are four types and each of
them is 25%

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F1 - dihybrid recessive parent
TtRr
× ttrr

RT Tr Tr Tr
tr TtRr Ttrr ttRr Ttrr

Phenotype or Genotype ratio of Dihybrid test cross = 1:1:1:1

 Construction : In test cross phenotypes and genotypes ratio are same.
 Trihybrid test cross (phenotype, genotype) ratio = 1 : 1 : 1 : 1 : 1 : 1 : 1 : 1
 Reciprocal Cross :
When two parents are used in two experiments in such a way that in one experiment ‘‘A’’ is used as the
female parent and ‘‘B’’ is used as the male parent, in the other experiment ‘‘A’’ will be used as the male
parent and ‘‘B’’ as the female parent. Such type of a set of two experiments is called Reciprocal cross.
Characters which are controlled by karyogene are not affected by Reciprocal cross. In case of cytoplasmic
inheritance result become change by Reciprocal cross.
(a) TT × tt (b) TT × tt
(Female) (Male) (Male) (Female)

All Tall All Tall

CYTOPLASMIC INHERITANCE (Correns)

Inheritance of characters which are contorlled by cytogene or cytoplasm is called cytoplosmic inheritance.
Genes which are present in cytoplasm called ‘Cytogene’ or ‘Plasmagene’ or extra nuclear gene. Total
cytogene present in cytoplasm is called ‘Plasmon’.
A gene which is located in the nucleus is called ‘karyogene’.
 Inheritance of cytogene in higher plants only through the female.
 The male gamete of higher plant is called male nucleus. It has very minute [equivelent to nil]
cytoplasm.So male gamete only inherited karyogene.
 Thus, inheritance of cytogene only through female cytoplasm. (also called maternal inheritance)
 If there is a reciprocal cross in this condition, then result will be effected.
Cytoplasmic inheretance of three types :
 Maternal effect depending indirectly on nuclear genes and involving no known cytoplasmic cytoplasmic
hereditary unit called as predetermination. maternal effect is determine before fertilization.
 Cytoplasmic inheritance involving dispensable and infective heredetary particle in cytoplasm which may
or may not depend on nuclear genes called as Dauermodification.
 Cytoplasmic inheritance involving essential organelles like, Chloroplast and mitochondria called as
organellar genetics.
Example of predetermination
Shell coiling in snail (Limnaea peregra)
In Shell coiling of snail can be dextral (Coiling to the right) or sinistral (coiling to the left). This direction
of coiling is genetically the dextral coiling depending upon maternal dominant allele D and sinistral
coiling depending upon maternal recessive allele (d).
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DD(O) dd(Q) DD(O)
+ dd(O)
+
P1 P2 P1 P2

dextral sinistral dextral sinistral

Dd Dd

F1 F1

dextral sinistral
Dd(O) Dd(Q) Dd(O)
+ + Dd(Q)

Inter Inter
cross cross
dextral dextral sinistral sinistral

DD Dd Dd dd DD Dd Dd dd

F2 F2

dextral dextral dextral dextral dextral dextral dextral dextral

F3 F3

dextral dextral dextral sinistral dextral dextral dextral sinistral

Above reciprocal cross indicates that phenotype of offspring is decided by genotype of female parent
not the phenotype of female parent. If female parent contains only one dominant gene ‘D’ then phenotype
of all offspring will be dextral.
Example :

Dd() dd() Dd() dd()

Sinistral dextral Sinistral dextral
F1 - All dextral F1 - All Sinistral

Example of Dauermodification -
 Sigma particle in Drosophila these particle are virus like particles which are present in Drosophila
and related to CO2 sensitivity. Inheritance of sigma particle takes place through the egg cytoplasm.
 Kaapa particle in Paramecium kappa particles are found in certain ‘‘Killer strains’’ of Paramecium
and are responsible for production of substance paramecin which is toxic to strain not prossessing
Kappa. (Sensitive Strain)
The minimum number of kappa particlesis 400 to secrete paramecin. Kappa particles are symbiotic
bacteria named ‘‘Caedobacter taeniospiralis’’
Example of Organellar Genetics : (True examles of cytoplasmic inheritance)
 Plastid inheritance in Mirabilis jalapa – cytoplasmic inheritance first discovered by Correns in
Mirabilis jalapa.
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 In Mirabilis jalapa plastid inheritance i.e. branch colour is example of cytoplasmic inheritance.
Branch colour
(a) Pale × Green (b) Green × Pale

F1 - Pale F1 - Green

(c) Varigated Pale or Green or Variegated

×

F1 - Pale, Green and Variegated

 Male strelity in maize plant – Gene of male sterelity present in mitochondria. If a normal male plant
crossed with a female plant which has genes of male sterelity then all the generation become male sterile
because a particular gene was present in female which inherited by female.
 Albinism in plant : Gene of albinism is found in chloroplast. Gene of albinism is leathal in plant.
 Inheritance of Bacterial plasmid. In bacteria plasmid inheritance is due to conjugation.
 Petite form in yeast = (mitochondrial gene) petite is mutant form of yeast. This mutant form is slow
growing on culture medium.
 Iojap inheritance in Maize Iojap is characterized by contrasting strip of green and white colour of leaves.
 Poky Neurospora - (Mitochondrial gene) poky is mutant form of Neurospora. It is slow growing on culture
medium.
Excerptions of conclusions of Mendel :
 Exception of Dominance : There are two exceptions of law of dominance. (i) Incomplete dominance;
(ii) Co-dominance,
 Incomplete Dominance : According to Mendel’s law of dominance. Dominant character must be
present in F1 generation. But in some organisms, F1 generation is different from the both parents.
Both factors such as dominant and recessive are present in incomplete dominance but dominant
factors are unable to express their character completely, resulting different type of generation is formed
which is different from the both parents.
Incomplete dominance was first discovered by Correns in Mirabilis jalapa. This plant is also called as
‘4 O’ clock plant ‘or’ Gul-e-Bans’.
G Three different types of plant are found in Mirabilis on the basis of flower colour, such as red, white
and pink. When plants with red flowers is crossed with white flower plants, pink flower obtained
in F1 generation the reason of this is that the genes of red colour incompletely dominant over the
genes of white colour.
When F1 generation of pink flower is self pollinated then the phenotypic ratio of F 2 generation is
red, pink, white = 1 : 2 : 1 in placed of normal monohybrid cross - 3 : 1
The ratio of phenotype and gentype of F2 generation in incomplete dominance is always same.

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Red × White
RR  rr
F1 geheration R r (Pink) 
 Self pollination

R r RR = Red - 1
R RR Rr
F2 – Generation Rr = Pink - 2
r Rr rr
rr = White - 1

G Antirrhinum majus - Incomplete dominance is also seen in this plant. This plant is also known
as ‘Snapdragon’ or ‘Dog flower’. Incomplete dominance is found in this plant which is the same
as Mirabilis flower colour.
G Andalusian Fowls - Incomplete dominance is present for their feather colour. When a black colour
fowl is crossed with a white colour fowl, the colour of F1 generation is blue.
 CO-DOMINANCE : In this phenomenon, both the gene expressed for a particular character in F1 hybrid
progeny.
Examples : Co-dominance is seen in animals for coat colour
When a black parent is crossed with white parent, a roan colour F 1 progeny is produced.
When we obtain F2 generation from the F 1 generation, the ratio of black; black white (Roan);white of
animals is 1 : 2 : 1
Note : F 2 generation is obtained in animals by sib-mating cross.
BLACK × WHITE
R1R1  R2R2
F1 generation R1R2 (Roan)
 Sib-mating cross
R1 R2
R1 R1R1 R1R2
R2 R1R2 R2R2
R1R1 = Black 1 R1R2 = Roan - 2 R2R2 = White - 1
It is obvious by above analysis that the ratio of phenotype as well as genotype is 1 : 2 : 1 in co-
dominance.
Sp. Note : In incomplete dominance, characters are blended phenotypically, while in co-dominace, both
the ganes of a pair exhibit both the characters and effect of both of both the character is independent
from each other.
Other Example of Co-dominance :
G AB blood group ibheritance (IAIB)
G Carrier of Sickle cell anaemia (Hb^ HbS)
 Conclusion of Segregation : There is no exception of law of segregation. The segeragation is essential
during the meiotic division in all sexually reproducing organisms. (Nondisiunction may be exception of the
law).

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 Exception of conclusion of independent assortment
The law of independent assortment is most criticised. Linkage is the exception of this law.
LINKAGE :
collective inheritance of character is called linkage first time seen by Bateson and Punnett in Lathyrus
odratus and gave coupling and repulsion phenomenon. But they did not explain the phenomenon of
linkage. Sex linkage was first discovered by Morgan in Drosophila & coined the term linkage. He proposed
the theory of linkage.
In 1906, Bateson and Punnet crossed two varieties of Lathyrus odoratus (sweet pea) and observed that
the results do not agree with the mendel’s law of independent assortment. They formulated the hypothesis
of coupling and repulsion to explain the unexpected F2 results of dihybrid cross between a homozygous
sweet pea having dominant alleles for blue flowers (BB) and long grains (LL) with another homozygous
double recessive plant with red flowers and round pollen grains (bbll).
Test Cross Ratio of F1 7 : 1 : 1 : 1 indicated that there was a tendency of the dominant alleles to remain
together. Similar was the case with recessive alleles.
Parents Blue flower & long pollen × Red flower & round pollen
BBLL bbll
Gametes BL × bl

F1 BbLl
Blue flower & Long pollen
Test cross BbLl × bbll
Test cross Progeny

7/16 : 1/16 : 1 / 16 : 7/16

Blue flower & Blue flower & Red flower & Red flower &
long pollen round pollen long pollen round pollen
(BbLl) (Bbll) (bbLl) (bbll)
It was called gametic coupling by bateson and Punnet. The tendency of two dominant genes to remain
together in the process of inheritance was called as coupling.
In another cross they took a sweet pea plant with blue flowers and round pollens (BBll) and other plant
with red flowers and long pollens (bbLL) and obtained the ratio of 1 : 7 : 7 : 1 by test crossing F 1
generation.
Parents Blue flower & round pollen × Red flower & long pollen
BBll  bbLL
Gametes Bl bL
F1 BbLl
Blue flower & long pollen
Test cross BbLl × bbll
Blue flower & long pollen  Red flower & round pollen

1/16 : 7/16 : 7/16 : 1/16

Blue flower & Blue flower & Red flower Red flower &
long pollen round pollen long pollen round pollen
(BbLl) (Bbll) (bbLl) (bbll)
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When two dominant or recessive genes comes from different parents, they tend to remain separate hence,
this ratio is called repulsion ratio. T.H. Morgan in 1910 showed that coupling and repulsion are two aspects
of the same phenomenon called linkage. He suggested that the two genes present on the same chromosome,
are in coupling phase and when present on two different homologous chromosomes are in repulsion
phase. Linkage therefore, may be defined as ‘‘The tendency of two genes of the same chromosome
to remain together in the process of inheritance’’.
According to Morgan, the degree or the strength of linkage depends upon the distance between the linked
genes in the chromosome.
Linkage and independent assortment can be represented in dihybrid plant
In case of linkage in dihybrid (AaBb) In case of independent assortment in
dihybrid (AaBb)
A a A a B b
B b

It produces two types of gamete It produces four types of gamete

AB : ab AB : ab : aB : Ab
Theory of linkage :
 Linkage genes are linearly located on same chromosome. They get separated if exchange (crossing over),
takes place between them.
 Strength of linkage  1 / distance between the genes. It means, if the distance between two genes is
increased then strength of linkage is reduced and it proves that greater is the distance between genes, the
greater the probability of their crossing over.
Crossing over obviously disturbs or degenerates linkage. Linked genes can be separated by crossing over.
Factors effecting crossing over (C.O.) & Linkage
G Distance  = C.O. Linkage 
G Temperature  = C.O. Linkage 
G X-Ray  = C.O. Linkage 
G Age  = C.O.  Linkage 

G Sex- Male C.O.  (Crossing over totally absent in male Drosophila) Linkage O > O

 Arrangement of linked Genes on Chromosomes :

The arrangement of linked genes in any dihybrid plant is two types.
G Cis - Arrangement : When, two dominant genes located on one chromosome and both recessive
genes located on another chromosome. Such type of arrangement is termed as cis-arrangement. Cis-
arrangement is an original arrangement.

Two type of gomets can be produced in this arrangement  AB and ab

G Trans arrangement : When a chromosome bears one dominant and one recessive gene, and another
chromosome also possess one dominant and one recessive gene, such type of arrangement is called
trans-arrangement. Trans arrangement is not an original form. It is due to crossing over. Two types of

gamete also formed in trans-arrangement but it is different from cis-arrangement Ab and aB .

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Cis Trans
A a A a

B b b B

AB ab Ab aB

Types of Linkage :
There are two types of linkage
 COMPLETE LINKAGE : Linkage in which genes always show parental combination. It never forms new
combination.
Crossing over is absent in it. Such genes are located very close on the chromosomes. Such type of linkage
very rare in nature e.g., male Drosophila, female silk moth.
 INCOMPLETE LINKAGE : When new combinations also appear along with parental combination in offsprings,
this type of linkage is called incomplete linkage, the new combinations form due to crossing over. The
percentage of new combination is equal to the percentage of crossing over. (<50%)
Example : In maize incomplete linkage was observed by Hutchinson. w.r.t seed coat colour and seed
shape.
The results show that parental combination of alleles (CS/CS and cs/cs) appear in about 96% cases. The
other two are new combinations (Cs/cs and cS/cs) apear in about 4% cases. Thus in about 4% cases
crossing over has occured between linked genes.
Parents Coloured & full × Colourless & shrunken
CS / CS cs / cs
Gametes CS × cs

F1 CS / cs
Coloured & full
Test cross Cs/cs × cs / cs
Gametes cs

Result : CS CS / cs – Coloured & full 48.2%

cs Cs / cs – Coloured & shrunken 1.8%

cS cS / cs – Colourless & full 1.8%

cs cs / cs – Colourless & shruken 48.2%

Incomplete Linkage
Another example was demonstrated by Morgan, while working with Drosphila. (i) Crossing of yellow bodied
(Y) and white eyed (W) female with brown bodied (Y+) red eyed (W +) male produced F1 to be brown bodied
red eyed. In F2 generation, obtained by selfing of F1 hybrids, the ratio deviated significantly from expected.
He found 98.7% to be parental and 1.3% as recombinants. (ii) In a second cross between white eyed and
miniature winged female (wwmm) wild red eyed normal winged male (W +W +M+M+) all hybrids were found
to be wild type. Test cross progency of this hybrid was found to be 62.8% parental and 37.2% recombinant.
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Linkag group : All the genes which which are loacated on one pair of homologous chromosome form one
linkage group. Genes which are located on homologous chromosomes are allelic so we consider one
linkage group.
 Linkage group – haploid no. of homologous chromosomes.
2n n Pair Linkage group
Human 46 23 23 23
Mouse 42 21 21 21
Frog 26 13 13 13
Maize 20 10 10 10
Pea 14 7 7 7
Drosophila 8 4 4 4
Neurospora 7 7 7 7
Bacteria / B.G.A. – – – 1
Application of Linkage :
Distance can be identified by the incomplete linakage. It’s unit is centi Morgan (cM).
1 1
Strength of linkage  Dis tance b / w linked gene  Cros sing Over

Genetic map / Linkage map / chromosome map – In genetic map different linked genes are linearly
arranged or chromosome according to percentage of crossing over ( Distance) between them.
With the help of genetic map we can find out the position of a particular gene on chromosome. Genetic
map is helpful in the study of genome.
Sex Linkage
When the genes of vegetative / somatic characters are present on sex-chromosome is termed as sex linked
gene and such phenomenon is known as sex-linkage. Two - types of sex linkage :
 X-linkage : Genes of sometic characters are found on x-chromosome. the inheritance of x-linked character
may be through the males and females.
e.g. Haemophilia, Colour blindness
 Y-linkage : The genes of somatic characters are located on Y-chromosome. The inheritance of such type
of character only through the males, such type of character is called Holandric character these characters
only found in male.
e.g.  Gene which forms TDF
 Hypertrichosis (excessive hair on ear pinna.)
Gene which is located on differential region of Y - chromosome is known as Holandric gene.
Example of X- linkage :
 Eye colour in Drosophila : Eye colour in Drosophila is controlled by a X-linked gene. If a red eyed
colour gene is represented as ‘+’ and white eyed colour represented as ‘w’, then on basis of this
different type of genotypes are found in Drosophila.
Gene for red eye domainant (+) and white colour of eye is recessive (w)
Homozygous red eyed female = X+X+
Heterozygous red eyed female = X+Xw
Homozygous white eyed female = XwXw
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+
Hemizygous red eyed male = X Y
Hemizygous white eyed male = XwY
It is clear by above different types of genotype that female either homozygous or heterozygous for eye
colour. But, for the male eye colour, it is always hemizygous.
 Haemophilia : Haemophilia is also called “bleeder’s disease” and first discovered by John Otto
(1803). The gene of haemophilia is recessive and x-linked lethal gene.
On the basis of x-linked, following types of genotype are found.
XhX = Carrier female
XhXh = Affected female
XhY = Affected male
But XhXh type of female dies during embryo stage because in homozygous condition, this gene is lethal
and causes death.
Haemophilia - A  due to lack of factor - VIII (Antihaemophilic globulin AHG)
Haemophilia B or Christmas disease - due to lack of factor - IX (plasma thromboplastin component)
 Haemophila -C - due to lack of factor XI (plasma thromboplastin anticedent)
 Colour Blindness : The inheritance of colour blindness is like as haemophilia, but it is not a lethal
disease so it is found in both male and female (discoverd by Horner)
Three types of colour blindness are :
G Protanopia : It is for red colour.
G Deuteranopia : It is for green colour
G Tritanopia : Blue colour blindness. Colour blindness is cheked by Ishihara - chart.

Other examples of X - linkage

 Diabetes insipidus (recessive).
 Duchenne muscular dystrophy (recessive)
 Fragile x syndrome (recessive).
 Pesudoricketes (Dominant)
 Defective enamel of teeth (Dominant)
Types of Inheritance of sex linked characters :
 Criss Cross inheritance (Morgan) : In criss-cross inheritance male or famale parent transfer a X-linked
character to grandson or grand daughter through the offspring of opposite sex.
G Diagenic : Inheritance in which characters are inherited from father to the daughter and from daughter
to grandson.
Father  Daughter  Grand Son
G Diandric : Inheritance in which characters are inherited from mother to the son and from son to grand
daughter. Mother  Son  Grand Daughter
 Non criss-cross inheritance : In this inheritance male or female parent transfer sex linked character to
grand son or grand daughter through the offspring of same sex.
G Hologenic : Mother  Daughter  Grand-daughter (female to female)
G Holandric : Father  Son  Grand-son (male to male)
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 Sex-Limited Character : These characters are expressed in one sex and unexpressed in another sex.
But their genes are present on autosome of both the sexes and their expression is depend on sex hormone.
Example : Secondary sexual characters  these genes located on the autosomes and these genes are
present in both male and female, but effect of these are depend upon presence or absence of sex-
hormones. For example - genes of beard & moustache express their effects only in the presence of male
hormone - testosterone.
 Sex Influenced Characters : Genes of these characters are also present on autosomes but they are
influenced differently in male and female. In heterozygous condition their effect is different in both the
sexes. Example : Baldness : Gene of baldness is dominant (B).

Genotype Male ( O ) Female ( O )

BB Baldness present Baldness present

bb Baldness absent Baldness absent
Bb Baldness present Baldness absent

Gene Bb shows partiality in male and female, Baldness is found in male, but baldness is absent in female
with this genotype.
MULTIPLE ALLELE
More than 2 alternative forms of a gene is called multiple allele.
Multiple allele is formed due to mutation.
Multiple allel located on same locus of homologous chromosome.
A diploid individual contains two allele and gamete contains one allele for a character.
Ex. Blood group - 3 alleles
Coat colour in rabit - 4 allels

n(n  1)
If n is the number of allele of a gene then number of different possible genotype =
2

Example of multiple allele :

 ABO blood group – ABO blood groups are determined by allele I . allele IB, allele I 0
IA = dominant
IB = dominant
IO = recessive
Possible phenotypes - A, B, AB, O
Blood Group Genotype Antigen or Antibody or
agglutinuogen agglutinin
A IAIA, IAI0 A b
B IBIB, IBIO B a
AB I^IB A & B None
O IOIO none a & b

3(3  1)
Possible genotype number = = 6 genotype
2


CAREERCoat
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Wild type = Full coloured = agouti = C+
Himalayan [white with black tip on extremities (like nose, tail and feet)] = c h
Chinchilla [mixed coloured and white hairs] = cch
albino = olourless = c a
These allels show order of dominance C+ > cch > ch > ca
Possible genotypes –
Coloured = C+C+, C+cch, C+ch C+ca
Chinchilla = cchcch, cchch, cchca
Himalayan = chch, chca
Albino = caca
4(4  1)
Possible genotype = = 10 genotypes
2
Eye colour in Drosophila and self incompatibility genes in plants are also the example of multiple
allelism.
LETHAL GENE :
Gene which causes death of individual in early stage when it comes in homozygous condition called lethal
gene. Lethal gane may be dominant or recessive both.
Many of these genes which do not cause definite lethelity are called semilethals.
In semilethal gene death occurs in late stage.
 Lethal gene was discovered by L.Cuenot in coat colour of mice, Yellow body colour (Y) was dominant
over brown colour (y).
Gene of yellow body colour is lethal.
So homozygous yellow mice are never obtained in population. It dies in embryonal stage.
When yellow mice were crossed among themselves segregation for yellow and brown body colour is
obtained in2 : 1 ratio.
Yy × Yy
Y y
Y YY Yy (Death)

y Yy yy
YY - death in embryonal stage so modified ratio = 2 : 1
 In plant lethal gene was first discovered by E.Baur in Snapdragon (Antirrhinum majus)
Golden leaves (G)
Snapdragon
Green leaves (g)
Golden × Golden
Gg Gg
G g
GG
G (Death) Gg

g Gg gg Modified Ratio : 2 : 1

Homozygous golden leaves are never obtained.

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 Sickle cell anaemia in human. In human, gene of sickle cell anaemia HbS is the example of sub
lethal gene. When two carrier individual of sickle cell anaemia are crossed then offsprings are obtianed.
HbSHbA × HbSHbA
HbS HbA
HbSHbS
HbS (Death) HbSHbA

Hb^ HbSHbA HbAHbA

Modified ratio 2 : 1

PLEIOTROPIC GENE :

Gene which controls more than one character is called pleiotropic gene.
This gene shows multiple phenotypic effect.
For example :

Seed coal colour

 In Pea plant : Single gene influences Red spot on leaf

Flower colour

 In Drosophila recessive gene of vestigial wings also influence the some another characters
G Structure of reproductive organs
G Longevity (Length of Body)
G Bristles on wings.
G Reduction in egg production.
 Example of pleiotropic gene in human.
G Sickle cell anaemia- Gene Hbs provide a classical example of pleiotrophy. It not only causes
haemolytic anaemia but also results increased resistance to one type of malaria that caused by
the parasite Plasmodium falciparum.
The sickle cell HbS allele also has pleiotropic effect on the development of many tissue and organs
such as bone, lungs, kidney, spleen, heart.
G Cystic fibrosis - Hereditary metabolic disorder that is controlled by a single autosomal recessive
gene. The gene specifies an enzyme that produces a unique glycoprotein.
This glycopotien results in the production of mucous.
More mucous interfere with normal functioning of several exocrine glands including those in the
skin, lungs liver and pancreas.
POLYGENIC INHERITANCE :
Inheritance of characters in which one character is controlled by many genes and intensity of character
depends upon the number of dominant allele or gene.
Polygenic inheritance first described by Nilsson - Ehle in kernal colour of wheat.
Nilsson - Ehle said explain kernal colour of wheat is regulated by two pairs of gene.

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RRBB × rrbb
Red White

F1- gen. RrBb (intermediate)

F2-gen. 1 : 4 : 6 : 4 : 1
Full light Intermediate e very white
red red red light
Number of 4 3 2 1 0
dominant allele
1 Red : 14 intermediate : 1 white
Example - Colour of the skin in Human.
The inheritance of colour of skin in human studied by Devenport.
When Negro (AA BB) phenotype crossed with white (aa bb) phenotype, intermediate phenotype produced
in F1 generation. Phenotypes of F2 generation as follows.

Negro × White
AA BB  aa bb
F1 - generation  Aa Bb (Intermediate)

F2

AB Ab a B ab
AB AA BB AA Bb AbBB AaBb
Negro Dark Dark Intermediate

Ab AA Bb AA bb AaBb Aabb
Dark Intermediate Intermediate Light

a B AaBB AaBb aa BB aa Bb
Dark Intermediate Intermediate Light

ab AaBb Aabb aaBb aabb

Intermediate Light Light White

Thus
Phenotypic ratio of F2 generation of quantitative inheritance as (2 pair)
Negro : Dark : Intermediate : Light : White
1 : 4 : 6 : 4 : 1
If human skin colour and kernal colour in wheat is regulated by 3 pairs of alleles so phenotypic
ration of F2 generation
Ratio 1 : 6 : 15 : 20 : 15 : 6 : 1
Negro Intermediate or Mullato white
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20/16

Frequencies of Individual
15/16

6/16

1/16

Intensity of colour
GENE INTERACTION :
Gene interaction is two types (i) allelic interaction : allelic interaction takes place between allele of same
gene which are present at same locus.
eg.  Incomplete dominance
 Co-dominance
Non allelic gene interaction : When interaction takes place between non allele is called non allelic gene
interaction. It changes or modifies other non allelic gene.
 Examples of nonallelic interaction :
G Epistasis : When, a gene prevents the expression of another non-allelic gene, then it is known as
epistatic gene and this phenomenon is known as Epistasis.
Gene which inhibit the expression of another non-alleleic gene is called epistatic gene and expression
of gene which is suppressed by epistatic gene is called hypostatic gene.
Example : Hair Colour in Dog :
B = Dominant allele for black colour of hairs
b = Recessive allele for brown colour of hairs.
I = Epistatic gene (White).
If the genotype bbii for brown colour and BBII for white colour. Following types of generation will be
obtained by following crosses.
bbll × BBII

F1 generation BbIi White colour
 F 2 generation

BI Bi bI bi
BI BBII BBIi BbII BbIi
White White White white
Bi BBIi BBii BbIi Bbii
White Black White Black
bI BbII BbIi bbII bbIi
White White White white
bi BbIi Bbii bbIi bbii
White Black White Brown

It is obviously clear by above analysis, the phenotypic ratio of F2 - generation in epistasis is -

12 : 3 : 1

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 Complementary Gene : Two pair of non allelic genes are essential in dominant form to produce a particuar
character.
Such genes that act together to produce an effect that neither can produce, its effect separately are called
complementary genes.
Both types of gene must be present in dominant form.
C – P purple coloured
C – pp colour less
cc – p colour less
cc – pp colour less
Row material Gene

C Gene P
 Chromogen   Anthocyanin (purple)
CC PP × cc pp
(coloured) (Colourless)

F1 - Generation  CcPp (Coloured)


CP Cp c P c p

CP CC PP CC, Pp Cc, PP Cc, Pp

Coloured Coloured Coloured Coloured

Cp CC Pp CC pp Cc, Pp Cc, pp
Coloured Colourless Coloured Colourless

c P Cc PP Cc Pp cc PP cc Pp
Coloured Coloured Colourless Colourless

c p Cc Pp Cc pp cc Pp cc pp
Coloured Colourless Colourless Colourless

Thus phenogypic ratio of complementary genes = Colourled : Colourless 9 : 7

MONOCLONAL ANTIBODY (MAB)

Cell. culture myeloma

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 Monoclonal antibodies are specific only for one antigen (or antigenic site) and synthesize outside the
animal body.
 Monoclonal antibody produced by a specialized cells through a technique called as hybridoma
technology.
 This technology was discovered by Georges Kohler and Milstein, were awarded with the 1984 nobel
prize.
 Each hybrid clone grown in culture medium to produce monclonal antibody.
 Monoclonal which acts as an enzyme called abzyme.

Examples of some autosomal characters in human

Character Dominant Recessive

Eye colour Brown / Black Blue

Eye size Large Smal
Hair Curly hair Straight
Cheek Dimple Normal
Hand Right Handed Left Handed
Rolling of tongue Roller Non-Roller
+
Rh Factor Rh Rh¯
PTC taster Taster Non Taster
Skin pigmentation Normal Albino

 CHROMOSOMAL THEORY OF INHERITANCE

This theory was proposed by Walter Sutton and Theodor Boveri (1902). Following are the main points
of the thory
 Male and Female gemetes play an equal role in contributing Parents T T t t
hereditary components of future generation.
 Gemetes serve as the bridge between two successive
generations. Gametes T T t t
 Only the nucleus of sperm combines with ovum. Thus, the
hereditary information is contained in the nucleus.
 Chromatin in the nucleus is associated with the cell division
F1 Generation T t
in the form of chromosomes.
 Any type of deletion or addition in the chromosomes can
cause structural and functional changes in living beings.
 A sort of parallelism is observed between Mendelian factors Gametes T t
and chromosomes.
 A number of genes or Mendelian factors are found on T T T t
chromosome. F2 Generation
 Determination of sex in most of the animals and plants is T t t t
affected by specific chromosomes. These chromosomes are
called sex chromosomes.
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Parallelism Between Gene and Chromosomes
 Chromosomes are also transferred from one generation to the next as in the case of genes (Mendelian
factors)
 The number of chromosomes is fixed in each living species. These are found as homologous pairs in
diploid cells. One chromosome from father and the other contributed by the mother constitute a
homologous pair.
 Before cell division, each chromosome as a whole and the alleles of genes get replicated and are
separated during mitotic division.
 Meiosis takes place during Gamete formation. Homologous chromosomes form synapses during
prophase-I stage which in later course get separated and transferred to daughter cells. Each gamete
or a haploid cell has only one allele of each gene present in the chromosome.
 A characteristic diploid number is again established by the union of the two heploid gametes.
 Both chromosomes and the alleles (Mendelian factors) behave in according to Mendel’s law of
segregation.
homologus chromosomes of a pure tall plant. allele (T) is found for tallness in each chromosome.
Likewise. in a pure dwrf plant (tt) allel (t) is present in each chromosome.
These homologous chromosome. Likewise. in a pure dwarf plant (tt) allele (t) is present in each
chromosome.
These homologous chromosomes get separated during meiotic division. Hence, each gamete possess
only one chromosoms each pair. Accordingly all the gamets of tall plants possess a chromosome with
an allele of tallness (T), while the gametes of dwarf plants possess a chromosome with an allele for
dwarfness (t). Their cross to produce F1 generation will yield tall hybrid plants with homologous
chromosomal pair containing Tt allele with homologous chromosomal pair containing Tt allelic pair. In
this generation two kinds of gamete will be formed during gametogenesis. 50% with the allele (T) for
tallness and 50% with the allele for dwrfness (t). Random combination of these gametes will produce
offsprings in F2 generation in the ratio of 25% pure tall (TT). 50% hybrid tall (Tt) and 25% dwarf (tt)

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SEX DETERMINATION
SEX DETERMINATION :
Establishment of sex through differential development in an individual at an early stage of life, is called sex
determination. Various methods operate in sex determination like enviromental, non-allosomic genetic
determination, allosomic sex determination and haplodiploidy
 Sex Determination on the basis of fertilization.
 Three types :
 Progamic : Sex is determined before fertilization.
eg. – Drone in honey bee
 Syngamic – Sex is determined during fertilization.
eg. – most of plants & animals
 Epigamic – Sex is determined after fertilization.
eg. – Female in honey bee.
Environmental Determination of Sex. It is non genetic determination of sex which is based purely
on environmental conditions. The organisms are potentially hermaphrodite and capable of expressing
any of the sexes.
 In marine worm Bonellia, larva develops into female if it settles down alone in an isolated place. Any
larva coming in contact with the already grown female, it changes into male, and lives as a parasite
in the uterus of female.
 Crepidula (marine mollusca) where larva develops into male in the company of female and develops
into female if left alone.
 In Crocodiles low temperature induces femaleness and high temperature maleness.
 In turtles temperature below 28ºC induces maleness, above 33ºC femaleness while between 28 - 33ºC
equal number of male and female animals are formed.
 In marine fish Medusa sex changes according to environmental condition, becoming male in cold water
and female in warm water.
 Allosomic determination of sex
Chromosomes are of two types :
 Autosomes or somatic chromosomes.
These regulate somatic characters.
 Allosomes or Hetersomes or Sex chromosomes
These chromosomes are associated with sex determination. Term “Allosome” & “Heterosome” were given
by Montgomery.
Sex chromosomes first discovered by “Mc Clung” in grass hopper
X - Chromosome discovered by “Henking” and called ‘x-body’.
Wilson & Stevens proposed chromosomal theory for sex determination.
 XX - XY type or Lygaeus type : This type of sex determination first observed by Wilson & Stevens in
Lygaeus insect. Two types.
 XX female and XY male : In this type of sex determination female is Homogametic produces one
type of gamete
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A+X
2A + XX (Female)  gametes
A+X
 Male is heterogametic (male produces two types of gamete)

A+X
2A + XY(Male)  gametes
A+X

In male X-chromosome containing gametes is called “Gynosperm” and Y-chromosome containing

gamete is called “Androsperm”
eg. Man and dioecious plants like Cocinea, Melandrium
 XY female and XX male or ZW female and ZZ male : In this type of sex determination female is
hetergametic produces two types of gamete and male individual is homogametic produces one type of
gamete.
It is found in some insects like butter flies, moths and vertebrates like birds, fishes and reptilies.
In plant kingdom this type of sex determination is found in Fragaria elatior.
 XX female and XO male : or “Protenor type” : In this type of sex determination deficiency of one
chromosome in male. In this type, female is homogametic and male is heterogametic.

A+X
Female (2A + XX) homogametic
A+X

A+O
Male (2A + XO) heterogametic
A+X

Example :
 Grass hopper
 Squash bug Anasa
 Cockroach
 Ascaris and in plants like - Dioscorea sinuta & Vallisneria spiralis
GENIC BALANCE THEORY :
C.B. Bridges proposed genic balance theory for sex determination in Drosophila
 According to Bridges in Drosophila Y-chromosome is heterchromatic so it have no role in sex
determination
In Drosophila sex determination takes place by sex index ratio.
No.of x chromosomes X
Sex index ratio = = (0.5 is male and 1 is female)
No. of set of Autosomes A
In Drosophila gene of femaleness (Sxl = Sex lethal gene) is located on x-chromosome and gene of
maleness is located on autosome
Gene of male fertility is located on y-chromosome and in Drosophila, y-chromosome plays addition role

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in spermatogenesis and development of male reproductive organ, so y-chromosome is essential for the
production of fertile male.

X
Sex index ratio  = 1  female (2A + XX), (3A + XXX)
A

X (2A + XY) = Fertile male

 = 0.5  male
A
(2A + XO) = Sterile male

X
 = 1.5  Super female or meta female (sterile) (2A + XXX)
A

X
 = less than 0.5  Super male or meta male (Sterile) (3A + XY)
A

X
 = In between 0.5 and 1  Intersex (Sterile) (3A + XX)
A

X 1
 = = 0.5 (Male)
A 2

Chromosmal diagram
of Drosophila
 Gynandromorph -
Body of some Drosophila has some cells with male (X0) and some cells with female genotype (XX). Body
of such type of Drosophila has half lateral part of male and half lateral part of female and it is called bilateral
gynandromorph. It is formed due to loss of one x-chromosome at metaphase plate during first zygotic
division. Formation of gynandromorph is the best evidence that y-chromosome does not play any role in
sex determination.

xo
XX

this X chromosome will be lost

HAPLOID - DIPLOID MECHANISM :
In insects of order Hymenoptera which includes ants. honey bees, wasps etc.
Sex determination takes place by sets of chromosomes.
Diploid (two sets)  Female
Haploid (One set)  Male
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In honey bee, male individual (Drone) develops from unfertilized eggs (Haploid.) Male is always parthenote.
Queen and worker bees develop from diploid eggs i.e. fertilized egg.

Feeds on Royal jelly

Queen
(Fertile female)
Fertilized egg  diploid larva

Worker
Bee bread (Sterile female)

SEX DETERMINATION BY HORMONE :

Dizygotic twins are common in cattle like cow, sheep, goat etc. Some times the placenta of the two
dizgytic twins fuse forming blood vascular connections between two developing foetus. If twins are dizygotic,
one foetus may be male and the other female.
 Male hormone produced before female hormone by male twins which suppress the differentiation
of female ( ) sex organ. Such a sterile female with underdeveloped overties ovducts, Uterous etc. is
called free martin.
In free martin conditions, female is sterile & male is normal.
Y spots :
Males are identified by Y spots (Zech 1970). Number of Y spots are equal to number of Y
chromosomes.
CYTOLOGICAL BASIS OF SEX DETERMINATION :
Barr body technique or Lyon’s hypothesis -
Interphasic nucleus of human female contains two X-chromosomes. Out of two, one X - Chromosome
becomes heterochromatin and other X - Chromosome is euchromatin. By staining X - heterochromatin, it
appears as a dens body which is called Barr body. (Facultative hetrochromatin)
No. of Barr body = (No. of X chromosomes - 1)
So a Normal female (2A + XX)  One Barr body
Normal male (2A + XY)  Barr body absent
Turner syndrome (Sterile female) (2A + XO)  No. Barr body
Klinefelter syndrome (Sterile male) (2A + XXY)  One Barr Body
Drum stick which occurs in blood of female of mammals, is also a type of barr body. Drum stick is absent
in neutrophils of Male.
SEX DETERMINATION IN HUMAN
There occur a special gene on differential region of Y-chromosome of human. called Sry - gene (Sex
determine region on y-chromosome). This gene forms a proteinaceous factor called TDF (testes determining
factor). TDF responsible for the development of male reproductive organs. So presence and absence of Y-
chromosome determines sex.
 Sex determination in plant :
H.E. Warmke discovered sex determination in Melandrium plant. In Melandrim Y- chromosome is long as
compare to X - chromosome. In plant sex chromosomes are found only in unisexual plant.
Pro. R.P. Roy gave the importance of Y-chromosome in plant.
He discovered sex determination in Coccinea Indica (Family - cucurbitaceae)
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Y - Chromosome contains four regions and X - chromosome contains two regions. Different functions of
these regions.
 Ist region - (Female suppressor region) : This region suppress the development of female reproductive
structures.
 IInd region (Male promotor region) : This region initiates or start the development of Anther
 IIIrd region (Male fertility region) : This region induces the further development of Anther.
 IVth region (Homologous region) : This region helps in the disjunction & Pairing of X and Y chromosome
during meiosis.
 Vth region (Differential region of X-chromosome) : This region induces the development of female gonads
So when one or more than one Y - chromosome is present then plant is male and in female plant Y-
chromosome is absent.

1 (female suppressor region)

2 (Male promotor region)

(Differential region of 5
X - chromosome) 3 (Male Fertility region)

(Homologous region) 4 4 (Homologous region)

X Y
 Special Case :
If Ist region of Y chromosome is removed then plant becomes bisexual
If IInd region of Y chromosome is removed then plant becomes female due to absence of IInd region, Ist region
of Y chromosome does not suppress the Vth region of X - chromosome.
If IIIrd region of Y chromosome is removed then plant becomes sterile male due to absence of III rd region
so further development of anther does not take place.
PHENOTYPIC EXPRESSION IN HAPLOID ORGANISMS
Diploid organisms such as pea and Drosophila, have two allels for each gene. With the result, the recessive
allele is not expressed in the phenotype in presence of the dominant one. However, this is not so in the
case of haploid organisms. Contrary to diploid organisms, the genetics of haploid organisms exhibit the
following features.
 Haploid organisms contain only one allele of a gene, so there is no complication of dominance. All the
genes, whether dominant or recessive, express itself in the offsprings.
 In absence of dominance, any new mutation is immediately expressed, in hapoid organism
 Study inheritance of mutated gene, linkage, crossing over and biochemical consequence of a mutation
can easily be studied in haploid.
Haploid parent A × a

Diploid zygote Aa

Meiosis

Haploid offspring A a

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LINKAGE AND RECOMBINATION IN NEUROSPORA (Drosophila of plant kindgom)
Detection of linkage and recombination of genes in haploid organisms as in fungi, bacteria etc. is
comparatively simple. Fungus Neurospora is one of the favourite material with geneticists, because :
 The life cycle is of a short duration.
 The life cycle of Neurospora is the product of a single meosis
 The meiotic products are linearly arranged in ascus as 8 ascospores as ordered tetrads (i.e., the
eight ascospores are arranged in the same order in which chromatids were on the meiotic metaphase
plate. This is called linear or ordered tetrad. Each of the four products of meiosis can be culutred
separately to study their phenotypes and genotypes. This is called tetrad analysis.

First meiotic division Second meiotic division meiotic

2n
diploid 4-haploid POM 8-ascospores
POM = Products of meiosis (Ordered Tetrad)
 First Division Segration Between Centromere and gene - a.
A cross between two strain of Neurospora, one normal (a+) and other mutant (a) strain produces 8-
ascospores, out of which four are normal (a+) and other four mutants (a). The linear arrangement of
ascospores in ascus is 4a+ : 4a. It indicates the absence of crossing over between locus-a and centromere.
This described as first division segration (4 : 4).
 Second Division Segregation Between Centromere and Gene-a
In a similar cross if crossing over takes place leading to paired arrangement of ascospores with a prticular
gene, it is described as second division segregation. The arrangement of ascopores in the sequence (2
: 2 : 2 : 2) is as follows :
a+ : a+ : a : a : a : a : a+ : a+ (2 : 4 : 2)
a : a : a+ : a+ : a+ : a+ : a : a (2 : 4 : 2)
a+ : a+ : a : a : a+ : a+ : a : a (2 : 2 : 2 : 2)

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 Single Gene Mapping in Neurspotra
In Neurospora centromere behaves as a gene for mapping gene pair. In such a case distance of gene from
the centromere is calculated by calculating the percentage of cross over between centromere and gene.
Que. If 10% asci show crossing over what will be distance between gene and centromere. If total 100 asci
are precsent in a Neurospora

90 non crossing over type

100
10 crossing over type

1 asci is derivative of 8 spores

100 asci are derivative of (100 × 8 = 800 spore)
10 asci are derivative of 80 spores
(Out of 80 only 40 will be the recombinant type)

recombinant spore
% C.O. = × 100
Total spore

40
= × 100 = 5% ans. 5 cM
800

GENE EXPRESSION
The second important characteristic (first is transmission) of the gene is to store and express the genetic
information that will contribute towards the phenotype.
DNA carries information for the synthesis of all proteins required for the function of a cell. A close
relationship between genes and enzymes (or gene control the metabolism) was first discovered by a British
physician Archibald Garrod in 1909 (Father of human biochemical genetics. Concept - One mutant gene
one metabolic block). He observed that certain hereditary diseases in man such as alkeptonuria (black
urine disease), albinism (absence of melanin pigment) phenylketonuria etc. are inborn errors of metabolism
and are due to the defect in the enyme that catalyses the conservation of one metabolic substance to
another. Thus, the inherited genetic defect is reflected in the deficiency of an enzyme.
Beadle and Tatum’s Hypothesis (One gene one enzyme hypothesis)
The concept that genes have the information to produce enzymes, or gene metabolism relationship was
experimentaly proved by Beadle and Tatum (1948), on the basis of experiments conducted on pink bread
mould (Neurospora crassa). This mould can normally grow in a simple minimal medium containing salts
and sugar, making all other chemicals such as amino acids, purines, pyrimidines etc. through enzyme
catalysed reactions. This wild type of the mould is called ‘protoroph’. Beadle and Tatum exposed the pink
bread mould to X-rays, which can bring about a change in the nucleotide sequence of DNA and thus causes
mutation. They found that the mutants were unable to grow on a minimal medium. Each type of mutant
required some extra nutrient in the minimal medium for it’s normal growth. Such nutritional mutants are
called ‘auxotrophs’. They obtained different nutritional mutants requiring amino acids ornithine or citrulline
or arginine for growth. The mutants could be classified into three types :
 Some could grow on ornithine -, or citrulline -, or arginine - containing medium,
 Some could grow on citrulline - or arginine containing medium,
 Some could grow on arginine supplemented medium
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Effect of medium supplemnts on the growth of mutants Neurospora crassa

Growth on Medium supplemented with

Mutant Type Ornithine Citrulline Arginine

I + + +
II – + +
III – – +
+ = growth – = No grwoth
Biochemical Pathway
Precursor Ornithine Citrulline Arginine

encoded enzyme Enzyme A Enzyme B Enzyme C

Location of gene Gene A Gene B Gene C

on chromosome
 This shows that all mutants could grow on arginine supplemented medium suggesting that arginine is
the final product of this pathway.
 Since some mutants could not grow on ornithine, this compounds must be synthesized earlier than
citrulline and / or arginine.
 Beadle and Tatum taking clue from this growth behaviour of Neursopora and other experiments, proposed
the arginine biosynthetic pathway. The precursor compound first gives rise to ornithine, ornithine
then gives rise to citrulline and the latter is finally converted into arginine; each of these steps is
mediated by an enzyme.
 The mutant of class I, could grow on ornithine, citrulline or arginine suggesting that they lack the
capacity to synthesize ornithine, but beyond that they could complete the pathway.
 Similarly, mutants of class II, could not convert ornithine to citrulline, but if supplied circulline could
Synthesize arginine.
 The last mutants of class III could not convert citrulline to arginine and had to be supplied Arginine
for growth.
 Beadle and Tatum reasoned that these defects could arise due to defective enzymes in each mutant.
Since such changes were mutational, they held that one gene controls one enzyme in a pathway
leading to their famous ‘One gene one enzyme hypothesis”.
The hypothesis states that each gene controls synthesis of a specific enzyme or protein. Beadle and
Tatum were awarded with Noble prize for this work in 1958. Their work founded the new science of
‘biochemical genetics’. Beadle and Tatum used total isolation method to detect the nutritional
mutant of Neurospora. Modification of Beadle and Tatum’s Hypothesis : One gene one enzyme
hypothesis held that a gene has information to produce one enzyme. The enzymes are proteins, but
all proteins are not enzymes. Recently some RNAs have also been found to mainfest enzyme activity.
Proteins are complex molecules and may be composed of one or more polypetide chains. For example,
haemoglobin consists of four polypetide chains - 2 and 2 chains. Thus, more than one gene may
control the synthesis of a protein. It is now held that one gene is responsible for the formation of one
polypetide chain. Therefore, one gene one enzyme hypothesis has been replaced by ‘one gene one
polypetide hypothesis’ Further modification came when one gene was identified as a functional unit
or cistron and the same was called ‘one cistron-one polypeptide hypothesis’.

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REGULATION OF GENE EXPRESSION :
 The mechanism which stimulates the expression of certain genes and inhibits other is called regulation
of gene expression.
 It is possible only if the organism has a mechanism of regulating gene activity by allowing some to
function and others to restrain their activity through switching on and switching off system. This means,
the genes are turned ‘on’ or ‘off’ as per requirement.
 A set of genes is ‘switched on’ when enzymes are required to metabolise a new substrate. The
enzymes produced by these genes metabolise the substrate.
 The molecules of metabolite the come to switch on of the genes are termed as inducers and the
phenomenon is called induction.
 Similarly, certain genes which are in their ‘switch on state, continue to synthesise a metabolite till the
later is produced in amount more than required or, it is supplied to the cell from outside. In other words
genes continue to express themselves till the end product inhibits or repress their expression Inhibition
by end product is known as ‘feed back repression’.
 OPERON CONCEPT :
 In 1961, two French microbiologist Francis Jocob and Jacques Monad at the Pasteur Institute in
paris, proposed a mechanism called operon model for the regulation of gene action in E. coli.
 An operon is a part of genetic matierial or DNA, which acts as a single regulated unit having one or
more structural genes-an operator gene, a promoter gene, a regulator gene.
 Openon is unit of Transcription.
 Operons are of two types (i) Inducible, (ii) Repressible.
G Inducible System (Lac operon of E. Coli)
 An inducible operon system normally remains in switched off condition and begins to work only when
the substance to be metabolised (Inducer) by it is present in the cell. Inducible operon system
generally occurs in catabolic pathways. e.g. Lac operon of E. coli.
Active repressor + inducer = inactive repressor

Galactose
Lactose
Glucose

An inducible opeon system consists of four types of genes

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 Structural genes : These genes synthesise mRNAs, which in turn synthesis polypeptide or enzyme
over the ribosomes. An operon may have one or more structural genes. Each structural gene of an
operon is called cistron. The lac operon (lactose operon) of Escherichia coli contains three structural
genes (z, y and a). These genes occur adjacent to each other and thus are linked. They transcribe
a polycistronic mRNA molecule, that helps in the synthesis of three enzymes - galactosidase or
Lactase, lactose permease and transacetylase.
 Operator gene : It lies adjacent to the structural genes and directly controls the synthesis of mRNA
over the structural genes. It is switched off by the presence of a repressor. An inducer can take away
the repressor and switch on the gene that directs the structural genes to transcribe.
 Promoter gene : This gene is the site for initial binding of RNA polymerase. When the operator gene
is turned on, the enzyme RNA polymerase moves over it and reaches the structural genes to perform
transcription.
 Regulator gene : It produces a repressor that binds to operator gene and stops the working of the
operator gene.
 Repressor : It is a protein, produced by the regulator gene. It binds to the operator gene so that the
transcription of structural gene stops. Repressor has two allosteric site (1) operator gene (2) effective
molecule (inducer / corepressor)
 Inducer : It is a chemical (substrate, hormone or some other metabolite) which after coming in contact
with the repressor, forms an inducer repressor complex. This complex cannot bind with the operator
gene, which is thus switched on.
The inducer for lac operon of Escherichia coli is lactose. When the sugar lactose is added to the culture
of E. coli, a few molecules of lactose gets into the bacterial cells by the action of the enzyme permease,
a small amount of this enzyme is present in the cell even when the operon is not working. These few
lactose molecules are then converted into an active form which acts as an inducer and binds to the
repressor protein. (allolactose an isomer of lactose) Allolacotse is real or true inducer of lac operon
The inducer repressor complex fails to join with the operator, which is turned on. The three genes are
expressed as three enzymes to metabolise lactose. Gratuitous inducer  Some molecules resemble with
natural inducer but are not metabolize by the enzyme. Example : Isopropyl this galactoside (IPTG) :
This resembles lactose and thus have the property of induction. Such inducer which induces enzyme
synthesis, without getting metabolized are called gratuitous inducer.
 Repressible System (Triptophan operon of E. coli)
A repressible operon system is normally in it’s switch on state and continue to synthesise a metabolite
till this metabolite is produced in amount more than required, or else it becomes available to the cell from
outside. Repressible operon system is commonly found in anabolic pathway. e.g. Tryptophan operon of
E.coli.
[Inactive repressor + co-repressor = active repressor]
Trytophan operon of Escherichia coli is an example of repressible system. It consists of the following:
 Structural operon These genes are meant for transcription of mRNA which in turn synthesis enzyme
:Tryptophan operon has five structural genes E.D.C. B and A. They lie in continuation and synthesis
enzymes for five steps of tryptophan synthesis.
 Operater gene (trp O) It lies adjacent to the structural genes and controls the functioning of the
structural genes. Normally, it is kept switched on, because the apo-repressor produced by the regulator
gene does not bind to it. The operator gene is switched off when a co-repressor is available alongwith
apo-repressor.

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 Promoter gene (trp P). It marks the site at which the RNA polymerase enzyme binds. When the
operator gene is switched on, it moes from promotor gene to structural genes for transcription.
 Regulator gene (trp R). It produces a regulatory protein called apo-repressor for (Inactive repressor)
blocking the activity of operator gene.
 Apo-repressor. (Inactive repressor) It is a regulatory protein synthesised by regulator gene. When a
co-repressor substance is available in the cell, the apo-repressor combines with the co-repressor to
form a apo-repressor co-repressor complex. This complex binds with the operator gene and switches
it off. Presence of apo-repressor alone, the operator gene is kept switched on because, by itself the
apo-repressor-is unable to back the working of operator gene.
Co-repressor – It is an end product of reaction by enzyme produced by the structure gens. In the presence
of tryphoton some molecules of tryptopham acts as co-repessor, co repressor bind with inactive repressor,
inactive repressor. co-repressor complex bind with operator region and prevent the binding of RNA
polymerase to the promoter, the trp-operon is off.

(i

Differeces between Induction and Repression

Induction Repression

1. It is switching on of an operon which 1. It is switching-off an operon which

is normally in switched off state (off-on) is normally in switched on state (on-off)
2. It starts transcription and translation. 2. It stops transcription and translation
3. It is caused by a new metabolite which 3. It is caused by incresed formation
needs enzymes to get metabolised. or availability of a metabolite (Feed back
repression).
4. It generally operates in a catabolic 4. It generally operates in an anabolic
pathway pathway

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The repressor molecules has key role in regulation of lac-operon. Repressor molecule active or
inactive. Active repressor may be inactive by addition of an inducer while the inactive repressor can be made
active by addition of a co-repressor.
Because the product of regulator gene the repressor act by shutting off the transcription of structural gene
the operon model, as originally proposed by Jocob & Monab is referred as negative control system.
 GENE EXPRESSION IN EUKARYOTES :
 Openon concept is not applicable for eukaryotes
 The most popular model is known as‘Britten-Davidson model’ or ‘Gene-battery model’ proposed by
Britten and Davidson in 1969.
 A set of structural genes controlled by one sensor site combindly called battery.
 Gene battery model assumes the presence of four class of sequences
G Producer gene : A producer gene is comparable to structural gene of prokaryotic operon.
G Receptor site : A receptor site is comparable to operater gene of bacterial operon and one such
receptor site is assumed to be present adjacent to each producer gene.
G Integrator gene : Integrator gene is comparable to regulator gene and is responsible for synthesis
of an activator RNA. It activates the receptor site.
G Sensor site : A sensor site regulates the activity of integrator gene. Activator gene can be transcribed
only when the sensor site is activated.
The sensor sites are recoginzed by agents which change the patterns of gene expression like hormones
and proteins. When a transcription factor (protein, hormone) bind to the sensor site it cause the transcription
of integrator.

TYPES OF GENE
All the genes do not play the same role nor all genes are active all the time. With regard to their role and
activity, the genes are of following types :
(1) Jumping Genes :
It is a segment of DNA which moves from one chromosome to another chromosome within the genome of
an indivisual. McClintock (1983) got nobel prize for the discoevery of jumping gene in maize. Two transposable
controling elements (Ds) and activator (Ac), which can jump to any chromosome from their original location
on chromosome 9. Also in bacteria plasmid transposone carry gene for antibiotic resistance (ampicilline).
Other Examples :
Transposable element (TE) in Drosophila - As much as 10% of the genome consist of transposons, most
important of these are copia like element, Fold back (FB) (for eye colour), and P and I element (for
sterlity). Ty elment in yeast.

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(2) Overlapping Gene : A few genes in certain bacteria and animal viruses code for two different polypetides.
These are called overlapping genes. For example – in  × 174 virus, SV -40 virus.
(3) Constitutive genes (House keeping genes):These genes are expressed constantly, because their products
are constant needed for cellular activity e.g. genes for glycolysis, gene of ATPase enzyme.
(4) Non-constitutive genes (Smart gene or Luxary gene) – These genes remain silent and are expressed
only when the gene product is needed. They are switched ‘on’ or ‘off’ according to the requirement of
cellular activities. Non-constitutive genes are of two types; inducible and repressible. The inducible
genes are switched on in presence of a chemical substance called inducer, required for the functioning
of gene activity. The repressible genes continue to express themselves till a chemical, often an end product
of the metabolism inhibits or represses their activity. Such type of inhibition is called feed back inhibition
or feed back repression.
(5) Homeotic genes : Homeotic gene regulates the organ differentiation in embryo.
 Homeobox : related to transcription of homeotic gene.
If mutation takes place in homoetic gene organ formation is disturbed.
(6) Pseudoallele : Allele which is located on non homologous chromosome or gene which is located on
different locus on homologous chromosome produces identical phenotype called as pseudoallele. eg
Duplicate gene
(7) Isoallele : If several allels exhibit same penotype then they are said to Isoallele. In Drosophila allele W +c
W +S W +9  produce red eye colour
(8) Hybrid vigour / Heterosis – Superiority of offsprings over it’s parents is called as Hybrid vigour or
Heterosis.
 Hybrid vigour can be maintained for long time in vegetaively propagated crops.
 Hybrid vigour can be lost by inbreeding (selfing) because inbreeding induces the Homozygosity and
reduced hetrozygocity in offsprings. Loss of Hybrid vigour due to inbreeding, is called as inbreeding
depression.
GENETIC ENGINEERING
INTRODUCTION :
 Gentic engineering also referred as ‘recombinant DNA technology’ or ‘gene splicing’ is one kind
of biotechnology involving manipulation of DNA
 It deals with the isolation of useful genes from a variety of sources and the formation of new combinations
of DNA (recombinant DNA) for repair, improvement, perfection and matching of a genotype.
 Thus genetic engineering may be defined ‘as a technique for artificial and deliberately modifying DNA
(gene) to suit human needs’.
 In genetic engineering manipulation breakage of DNA molecule of two desired places with the help of
rectriction endonuclease to isolate a specific DNA segment and then insert it in another DNA molecule
at a desired position.
 The new DNA molecule is recombinant DNA and the technique called genetic engineering. Genetic
engineering aims at adding. removing or repairing of a part of genetic material. Genetic engineering can
be used to improve the quality of human life.
The construction of the first recombinant DNA emerged from the possibility of linking a gene encoding
antibiotic resistance with a native plasmid (autonomously replicating circular extra chromosomal DNA)
of Salmonella typhimurium. Stanley Cohen and Herbert Boyer accomplished this in 1972 by isolating
the antibiotic resistance gene by cutting out of piece of DNA from a plasmid which was responsible
for conferrring antibiotic resistance. The cutting of DNA at specific locations became possible with the
discovery of the so called ‘molecular scissors’ - restriction enzymes. The cut piece of DNA was then
linked with the plasmid DNA. These plasmid DNA act as vectors to transfer the piece of DNA attached
to it.
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Paul berg (Father of genetic engineering). He transferred gene of SV 40 virus (simian virus) in to
E.coli with the help of  – phage. (Nobel prize - 1980)
The concept of genetic engineering was the outcome of two very significant discoveries made in
bacterial research. These were
G presence of extrachromosomal DNA fragments called plasmids (Magic circle) in the bacterial cell,
which replicate along with chromosomal DNA of the bacterium.
G presence of enzymes restriction endonucleases which cut DNA at specific sites. These enzymes
are, therefore, called ‘molecular scissors’
 TOOLS AND TECHNIQUES OF GENETIC ENGINEERING
Tools :
Genetic engineering involves cutting of desired segments of DNA and pairing of D.N.A in a vector to produce
a recombinant DNA (rDNA). The ‘biological tools’ used in the synthesis of recombinant DNA include
enzymes, vehicle or vector DNA, passenger DNA and alkaline phosphatases.
(1) ENZYME
 Lysing enzymes : These enzymes are used for opening the cells to get DNA for genetic experiment.
Bacterial cell wall is commonly dissolved with the help of lysozyme.
 Cleaving enzymes : These enzymes are used for DNA molecules. Cleaving enzymes are of three
types; exonucleases, endonucleases and restriction endonucleases.

G Exonucleases cut off nucleotides from 5’ or 3’ ends of DNA molecule.

G Endonucleases break DNA duplex at any point except the end.
G Restriction endonucleases cleave DNA duplex at specific points in such a way that they come to
posses short single stranded free ends. For example, a restriction endonuclease ECOR-I (from
Escherichia coli) recoginzes the base sequence GAATTC / CTTAAG in DNA duplex and cleaves
it’s strands between G and A.
Restriction enzymes are obtained from bacteria. They are useful to bacteria because the enzyme bring
about fragmentation of viral DNA without affecting the bacterial genome. This is an protective adaptation
against baceriophages. Restriction enzyme (Eco R - I) was discovered by Arber, Smith & Nathans
(1978 Nobel prize). These enzymes exist in many bacteria beside cleavage some restriction
endonuclease, also have capability of modification. Modification in the form of methylation, by methylation
the bacterial DNA modifies own DNA and therofore protects its own chromosomal DNA. Restriction
enzymes are used in recombinant DNA technology because they can be used in vitro to recoginze and
cleave within specific DNA sequence typically consisting of 4 to 8 nucleotides. The specific 4 to 8
nucleotide sequence is called restriction site and is usually palindromic, this means that the DNA
sequence is the same when read in a 5’-3’ direction on both DNA strand

AND MADAM DNA

As a results the DNA fregments produced by cleavage with these enzymes have short single stranded
overhang at each these kinds of ends are called sticky or cohesive ends because base pairing between
them can stick the DNA molecule back together again.
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5 G A A T T C 3' 5 G 3' 5 A A T T C 3'

 +
3'C T T A A G 5' 3'C T T A A 5' 3' G 5'

Therefore by cutting two different DNA samples with the same restriction enzyme and mixing the
fragments together a recombinant DNA molecule can be generated.
Exceptionally, some enzymes cleave both strand of DNA at exactly the same nucleotide position,
typically in the center of the recognition sequence resulting in blunt end or flush end.
Sma I (Serratia marcescens)

5' C C C G G G 3' 5' C C C 3' + 5' G G G 3'


3' G G G C C C 5' 3' G G G 5' + 3' C C C 3'

Nomenclature of enzyme : The first letter used for the enzyme is the first letter of the bacterium
genus name (in Italics) then comes the first two letter of it’s species (In Italics), next is the strain of
the bacteria, last is Roman numerical signifing the order of discovery of Bacteria.

EXAMPLES OF RESTRICTION ENZYME

Recognition sequences of some restriction endonucleases

Name Recognition End after cleavage Source

sequence

Eco RI  Escherichia coli - containing

– GAA T T C – –G AAT T C – drug resistant plasmid RI
– C T TAA G – – C T TAA G–

Hind III  –A AGCTT– Haemophilus influenzae
–AA G C T T – – TTC GA A–
– T T C GAA –

Bam I  –G GAT C C – Bacillus amyloliquefaciens
– G GATC C – – CCTAG G–
– CCTAG G –

Hae III  –GG CC– Haemophilus aegyptius
–GGCC– –CC GG–
–CCGG–


 Synthesizing enzymes : These enzyme are used to synthesize new trands of DNA, complementary
to existing DNA or RNA template. They are of two types; reverse transcriptases and DNA polymerases.
G Reverse transcriptases help in the synthesis of complementary DNA strands on RNA templetes.
G DNA polymerases help in the synthesis of complementary DNA strands on DNA tempates

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 Joining enzymes : These enzymes help in joining the DNA fragments. For example DNA ligase from
Escherichia coli is used to join DNA fragments. Joining enzymes are, therefore, called molecular glues.
 Alkaline phosphatases : These enzymes cut off phosphate group from the 5’ end of linearised circular
DNA (plasmid) and prevent its recircularisation.
(2) PASSENGER DNA :
It is the DNA which is transferred from one organism into another by comining it with the vehicle DNA. The
passenger DNA can be complementary, synthetic or random.
 Complementary DNA (cDNA) : It is synthesized on mRNA template with the help of reverse
transcriptase and necessary nucleotides. The DNA strand is then separated from the hybrid RNA-DNA
complex by using alkali. Complementary DNA strand is then synthesized over the template of DNA with
the help of DNA polymerase. cDNA formed through reverse transcription is shorter than the actual or
in vivo gene or DNA because of the absence of introns or non-coding regions.
 Synthetic DNA (s-DNA) : It is synthesized with the help of DNA polymerase on DNA template.
Kornberg (1961) synthesized first synthetic DNA from a mixture of deoxyrinonuclotide triphophates
DNA polymerase enzyme, metal ion and a segment of viral DNA Khorana (1968) synthesized first
artificial gene (DNA) without a template. They syntesized the gene coding for yeast alanine t-RNA,
which contained only 77 base pairs. However, it did not function in the living system. In 1979, Khorana
was able to synthesize a functional tyrosine t-RNA gene of E.coli with 207 nucleotide pairs. Since then
a number of genes have been synthesized artifically.
 Random DNA - It refers to small fragments formed by breaking a chromosome with the help of
restriction endonucleases.
(3) VEHICLE DNA OR VECTOR DNA :
The DNA used as carrier for transferring a fragment of foreign DNA into a suitable host is called vehicle
or vector DNA.
You know that plasmids and bacteriophages have the ability to replicate within bacterial cells independent
of the control of chromosomal DNA.
 CHARACTERISTIC OF VECTOR
 Origin of replication (ori) : This is a sequence from where replication starts and any piece of DNA
when linked to this sequence DNA can be made to replicate within
the host cells. This sequence is also responsible for controlling the
copy number of the linked DNA. So, if one wants to recover many
copies of the target DNA or gene it should be cloned in a vector
where origin support high copy number.
 Selectable marker : In addition to ‘ori’, vector requires a selectable
marker. Normally, the genes encoding resistance to antibiotics such
as ampicilin (ampR), chloramphenicol, tetracycline (tetR) or
kanamycin, etc., are considered useful selectable markers for E. coli.
 Cloning sites : In order to link the alien DNA, the vector needs,
recognition sites for the commonly used restriction enzymes. The ligation of alien DNA is carried out
at a restriction site present in one of the two antibiotic resistance genes. For example, you can ligate
a foreign DNA at the Bam H 1 site of tetracycline resistance gene in the vecter pBR322. The recombinant
plasmids will loss tetracycline resistance due to insertion of foreign DNA (insertional inactivation) now,
it can be selectd out from non-transformant ones by plating on ampicillin containing medium. The
transformants (plasmid transfer) growing on ampicillin (Bacteriolytic) containing medium are then
transferred on a medium containing tetracycllin (Bacteriostatic). The recombinants will not grow on the
medium containing tetracyclic antibiotics (due to insertion of inactivation). In this case, one antibiotic
resistance gene helps in selecting the transformants and another help in selecting recombinant.
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Selection of recombinants due to inactivation of antibiotics is a cumbersome (troublesome) procedure
because it requires simultaneous plating on two plated having different antibiotics. Therefore, alternative
selectable markers have been developed which differentiate recombinants from non-recombinants on the
basis of their ability to produce colour in the presence of a chromogenic substrate. In this, a recombinant
DNA is inserted within the coding sequence of an enzyme, which is referred to as insertional inactivation.
The presence of a chromogenic substrate X - gal (5 –bromo–4–chloro––D galacto pyranoside) gives blue
coloured colonies if the plasmid in the bacteria does not have an insert or presence of insert recombinant
results into insertional inactivation of the -galactosidase (reporter enzyme) and the colonies do not
produce any colour, these are identified as recombinant colonies.
 Vectors for cloning genes in plants and animals : Agrobacterium tumifaciens, a pathogen of several
dicot plant deliver a piece of DNA known as ‘T-DNA’ to transformer normal plant cells into a tumour.
Similarly, retroviruses in animals have the ability to transform normal cells into cancerous cells. better
understanding of the art of delivering genes by pathogens in their eucaryotic hosts has generated knowledge
to transform these tools of pathogens into useful vectors for delivering genes of interest to human. The
tumour inducing (Ti) plasmid of Agrobacterium tumifaciens has now been modified into a cloning vector.
 Plasmids : They are extra chromosomal DNA segments found in bacteria which can replicate
indepndently. Plasmids can be taken out of bacteria and made to combine with descried DNA segments
by means of restriction enzymes and DNA ligase. A plasmid carrying DNA of another organism integrated
with it, is known as reombinant plasmid or hybrid plasmid or Chimeric plasmid.
 Viruses : The DNA of cenrtain viruses is also suitable for use as a vehicle DNA. Bacteriophage
(bacterial virus) has been used to tranfer gene for  galactosidase from Escherichia coli to human cells.
Lambda phage ( phage) has been used for transferring lac genes of E. coli into haplid callus of tomato.

Vactor type Insert size kb (Kilobase)

Plasmid 0.5 – 8
Bacterophage lambda 9 – 23
Cosmid 30 – 45
BAC 50 – 300
YAC 1000 – 2500
 Technique of Recombinant DNA
 The DNAs which are to be used as passenger DNA and the vehicle DNA are extracted out of their cells
by lysing the cells with the suitable enzyme.
 The extracted DNAs are isolated from other cell contents by ultra centrifugation and purified.
 Both the passenger and vehicle DNAs are then. cleaved by using the same restriction endonuclease
so that they have complementary sticky ends.
 The complemetry sticky end of the passenger and vehical DNA are joined with ligase enzyme. This
gives its to a combined DNA
 The recombinant DNA is now inserted into a host cell such as Escherichia coli. The bacteria to be used
as hosts should be without plasmids.
 The host cells are treated with calcium chloride or lysozyme. It creates transient (temporary) pores in
their wall and makes the latter permeable to recombinant DNA.
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Escherichia coli Human cell

Plasmid Bacterial
Chromosome
Restriction
Enzyme
insulin gene

Sticky ends
Cut plasmid
Annealing by
DNA ligase

Plasmid free E. coli

Recombinant DNA

Multiplication of
recombinant DNA
reintroduced

Clones of engineered
bacteria medium

Recombinant insulin

STEPS INVOLVED IN GENE TRANSFER FOR THE

PRODUCTION OF HUMAN INSULIN

 The recombinant DNA is added to the culture in which such bacteria are growing. The recombinant DNA
is taken up by the bacteria. It replicates when the host bacteria divide and give rise to multiple copies
of recombinant DNA.
 GENE TRANSFER
Transfer of desired genes from one organism into another is an important aspect of genetic engineering.
Gene transfere involves essentialy three stages.
 Isolating a useful DNA segment from the donor organism.
 Inserting it to a suitable vector.
 Splicing of this altered DNAs into a recipient organism or host cell.
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The cell to begin making the appropriate product and then device an economical method for it’s mass
production.
Gene transfer is achieved by two kinds of transfer methods. (i) indirect method throughvector or
carriers and (ii) direct or vector less transfer method.
 Gene Transfer Through Vectors : Plasmids and viruses are commonly used vectors for the transfer of
desired genes. The desired genes are first made to join suitable plasmid or virus which are then introduced
into the target cells.

G A plant pathogenic bacterium -Agrobacterium tumefaciens produces crown galls or plant tumours in
almost all dicotyledonous plants.
G This bacterium infects all broad leaved agricultural crops such as tomato, soyabean, sunflower and
cotton but not cereales.
G Tumour formation is induced by it’s plasmid which is therefore called Ti plasmid (Ti for tumour inducing)
Agrobacterium tumefaciens naturally transfers some part of Ti-plasmid in to host plant DNA without any
human effort so it is called natural genetic engineer of plant.
In the transformation process two essential component in Ti - plasmid -
G T - DNA – (Transferred DNA)
G Vir-region – (Virulence region)
Insider the host plant cell T-DNA is seperated from Ti plasmid, and integrated into host plant DNA that
causes crown gall tumour.
Vir-region contains genes which are essential for T-DNA transfer and integration to host plant DNA. Vir-
region contains 6-operons A, B, C, D, E and G (Acetosyringone is inducer of Vir-operon)
When we use Ti plasmid as a vector, first we remove the tumour causing gene from T-DNA region. Then
desired gene inserted inplace of it. Now, this plasmid is called disarmed plasmid.
Same as Ri plasmid of A.rhizogenes (causing hairy root disease) also used as vector for gene transfer
to plant cell.
 Vectorless Gene Transfer
Foreign genes can also be transferred directly by the following methods.
 Electroporation : It creates transients (temporary pores) in the plasma membrane to faciliate entry
of foregin DNA.
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 Chemical mediated genetic transformation : It involves certain chemicals such as polyethylene
glycol (PEG), that help in the uptake of foregin DNA into host cells.
 Microinjection : It is the introducion of foreign genes into plant or animal cells using micropipettes
or glass needles.
 Particle gun : It is a technique in which tungsten or gold particles coated with foreign DNA are
bombarded into target cells to faciliate entry of the foreign genes.
 Liposome mediatd gene transfer : In this method DNA encloses within lipid begs. These lipid begs
fused with protoplast.
 GENE TRANSFER IN ANIMALS
In animals, the genes are transferred mostly through direct methods such as electroporation, microinjection
or using particle gun.
The desired foregin genes can be intorduced into fertilised eggs or embroys through microinjection. These
transgenic eggs or embroyos can be implanted into the uterus of another female, called surrogate
(foster)mother for their further development. Now, since most of the human genes have been identified
through ‘Human Genome Project’, It is hoped that a number of human genetic disorders such as
Alzheimer, cancer, haemophilia, thalaessemia and cystic fibrosis can now be cured through of the correct
genes into these patients.
 ACHIEVEMENTS OF GENETIC ENGINEERING :
 The DNA recombinant technology or genetic engineering provides great benefits for advancement of science
and society
 A new system of medicine gene therapy, may develop to treat hereditary diseases such as haemophilia.
G Genetic disorder can be over come by intorducing specific gene.
 Bacteria may be used as ‘Living factorries’ for synthesizing vitamins, hormones and antibodies.
G Human insulin (Humulin) was first genetically engineered product produced by an american firm Eli
Lilly – 5th July 1983.
G Charles weismann of university of Zurich, obtained interferon through recombinant E.coli (1980)
G Microbes have been engineered to produce Human growth Hormone (HGH) for curing dwarfism.
G Vaccines which are produced by genetic engineering e.g., for Hepatitis B and Herpes virus.
G Nitrogen fixation gene may tetrasferred from bacteria to major food crops to boost food production
withou using expensive fertilizers.
G Transgenic plant obtained through recombinant DNA technology. First transgenic plant was tobacco.
It contains resistant gene against weedicide (Glycophosate).
First transgenic animal was mouse contain gene for growth hormone.
First introduced transgenic crop in India (2002) is Bt-cotton.
It is resistant for ball worm (Helicoperpa armigera - Larva of insect). It is formed by transfer of pest
resistant gene from Bacillus thuringiensis (bt-2 gene encoding Bt-toxin).
Bacilus thuringiensis produces a protoxic protein called crystal protein (Cry-Protein) this protein is toxic
for larva of certain insect. This protein is converted to toxin by protease in gut this active toxin cause
lysis and poration of gut wall
G In pollution control, microbes have been engineered to break up the crude oil spills.
Dr. Ananda Mohan Chakraborthi Introduced plasmid from different strains in to single cell of
pseudomonas putida. The result was new genetically engineered bacterium which would cleaning oil
spills called ‘‘Super bug’’ (oil eating bug)

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Applications of ecombinant DNA products

Medically useful Applications

recombinant products

Human insulin Treatment of insulin - dependent diabetes

Human growth homone Replacement of missing hormone in short stature people.
Calietonin Treatment of rickets
Chorionic gonadotropin Treatment of infertility.
Blood clotting factor VIII / IX Replacement of clotting factor missing in patients with
Haemophilla A/B
Tissue Plasminogen activator (TPA) Dissolving of blood clots after heart attacks and strokes.
Erythropoetin Stimulation of the formation of erythrocytes (RBCs) for
patients suffering from anaemia during dialysis or side
effects of AIDS pattents treated by drugs.
platelet derived growth factor Stimulation of wound healing
Interferon Treatment of pathogenic viral infections, cancer
Interleukin Enhancement of action of immune system
Vaccines Prevention of infectious diseases such as hepatitis
B, herpes, influenza, partusis, meningitis, etc.

Application of Genetically Engineered Microbes

Microbes Applications

Escherichia coli Production of human insulin, human growth factor

Interferons, interleukin and so on.
Bacillus thuringiensis (soil bacterium) Productions of endotoxin (Bt toxin), highly potent, safe
and biodegradable insecticide for plant protection.
Rhizobium meliloti (bacterium) Nitrogen fixatin by incorporating ‘‘nif’’ gene in cereal crops
Pseudomonas putida (bacterium) Scavenging of oil spills by digesting hydrocarbons of crude
oil
Bacterial strains capable of Bioremediation (cleaning of pollutants in the enviroment).
accumulating heavy metal
Trichoderma (fungus) Production of enzyme chitinases for biocontrol of fungal
diseases in plants.

Transgenics and their potential applications

Transgenic Useful appications

Bt Cotton Pest resistance, herbicide tolerance, and high yield
Flavr Savr Tomato Increased shelf-life (delayed ripening) and better nutrient
quality
Golden Rice Vitamin A and Fe - rich
Cattles (cow, sheep, goat) Therapeutic human proteins in their milk
Pig Organ transplantation without risk of rejection
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Medical Diagnosis of Disease :
Recombinant DNA technology is one of the important tool for the diagnosis of several diseases. The
diagnositic technique involves the construction of probes consisting of short segments of single stranded
DNA attached to a radioactive or fluoroscent marker. Such probes are used to identify infections agents
such as Solmonella (cause food poisoning). Staphylococus (pus forming bacterium), hepatitis virus, HIV;
muscular dystrophy, cystic fibrosis and so on. Recombinant DNA technology can also be employed to
predict the inheritance of genetic disorders from carrier parents. The chances of birth of an affected child
can be predicated by testing DNA of repetitive prospective genetic disorder carrier parents.
 GENE THERAPY
 Techinique for curing genetic disease through replacing a ‘‘Faulty Gene’’ by a normal healthy functional
gene.
 The first gene therapy used in severe combined immuno deficiency (SCID) pateint.
 ADA enzyme involved in purine metabolism.
 These patients have no functioning T & B lymphocytes.
 The affected child of SCID develops recurrent infection early in life because they have no immune
response against invading pathogen.
 The ideal approach would be to give the patient a functioning ADA gene by gene therapy.
DNA FINGER PRINTING / DNA TYPING / DNA PROFILING / DNA TEST

 It is technique to identify a person on the basis of his/her DNA specificity. This technique was invented
by sir Alec. Jeffery (1984).
 In India DNA finger printing has been started by Dr. V.K. Kashyap & Dr. Lal Ji Singh.
 DNA of human is almost the same for all individuals but very small amount that differs from person to
person that forensic scientists analyze to identify people.
These differences are called Polymorphism (many forms) and are the key of DNA typing. Polymorphisms
are most useful to forensic scientist. It is consist of variation in the length of DNA at specific loci is
called Restricted fragment. It is most important segment for DNA test made up of short repetitive
nucleotids sequences, these are called VNTRs also called minisatellites. Restricted fragement consist
of hypervariable repeat region of DNA having a basic repeat sequence of 11-60 bp and flanked on both
sites by restriction site.
 The number and position of minisatellites or VNTR in restriction fragment is different for each DNA and
length of restricted fragment is depend on number and pattern of VNTR.
 Therefore, when the genome of two people are cut using the same restriction enzyme the length of
fragments obtained is different for both the people.
 These variations in length of restricted fragment is called RFLP or Restriction fragment length
polymorphism.
 Restriction Fragment Length Polymorphism distributed throughout human genomes are useful for DNA
Finger printing.
 DNA Fingerprint can be prepared from extremely minute amount of blood, semen, hair bulb or any other
cell of the body.
DNA content of 1 - Microgram or 100,000 cell is sufficient.

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Technique of DNA Finger printing involves the following major stpes.
 Extraction - DNA extracted from the cell by cell lysis. If the content of DNA is limited then DNA can
be amplified by Polymerase chain reaction (PCR). This process is amplification.
RESTRICTION
ENZYME
DIGESTION

GEL ELECTROPHORESIS
(Agargose polymer ge)

DENATURATION BY ALKALI SOLUTION

SOUTHERN BLOTTING

NYLON OR NITROCELLULOSE SHEET

HYBRIDIZATION

DNA PROBES

 Restriction Enzyme Digestion : Restriction enzyme cuts DNA at specific 4 or 6 base pair sequences
called restriction site.
Hae III (Haemophilus aegyptius) is most commonly used enzyme. It cuts the DNA, every where the
bases are arranged in the sequence GGCC. These restricated fragment transferred to Agarose Polymer
gel.
 Gel Electrophoresis :
 Gel electrophoresis is a method that separates macromolecules-either nucleic acid or proteins on
the basis of size, electric charge.
 A gel is colloid in a solid form. The term electrophoresis describes the migration of charged
particles under the influence of an electric field. Electro refers to the energy of electricity. Phoresis,
from the Greek verb phros, means “to carry across.” Thus, gel electrophoresis refer to the technique
in which molecules are forced across a span of gel, motivated by an electrical current. Activated
electrodes at either end of the gel provides the driving force. A molecule’s properties determine, how
rapidly an electric field can move the molecule through a gelatinous medium.
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 Many important bilogical molecules such as amino acids, peptides, proteins, nucleotides, and
nucleic acids posses ionisable groups and, therefore, at any given pH, exist in solution as electrically
changed species either as cation (+) or anions (–) Depending on the nature of the net charged
particles will migrate either to the cathode or to the anode.
 By the gel electroporesis these restricted fragments move towards the positive electrode (anode)
because DNA has –ve electric charge (PO4–3).
 Smaller Fragment more move towards the positive pole due to less molecular weight. So
after the gel electrophoresis DNA fragment arranged according to molecular weight or
length.
 These separated fragments can be visualized by stanining them with a dye that fluoresces ultraviolet
radiation.
 This appears the specific restricted fragment length pattern. The length pattern is different in
different individual.
This is called Restricted Fragment length Polymorphism (RFLP).
 Southern tranfer / Southern blotting :
The gel is fragile. It is necessary to remove the DNA from the gel and permanently attaches it to a solid
support. This is accomplished by the process of Southern blotting. The first step is to denature the DNA
in the gel which means that the double stranded restriction fragments are chemically separated into the
single stranded form. The DNA then is transferred by the process of blotting to a sheet of nylon. The nylon
acts like an ink blotter and “blots” up the separated DNA fragments, the restriction fragments, invisible at
this stage are irreversible attached to the nylon membrane the “blot”.
This process is called Southem blot by the name of Edward Southern (1970).
 Hybridization : To detect VNTR locus on restricted fragment, we use single stranded Radioactive (P32)
DNA probe which have the base pair sequences complimentary to the DNA sequences at the VNTR locus.
 Autoradiography : Nylon membrane containing radio active probe exposed to X-ray. Specific bands appear
on X-ray film. These bands are the areas where the radioactive probe bind with the VNTR.
These allow analyzer to identify a particular person DNA. The occurance and frequency of a particular
genetic pattern contained in this X-ray film. These x-ray film called DNA signature of a person which is
specific for each individual.
The probability of two unrelated individual having same pattern of location and repeat number of minisatellite
(VNTR) is one in ten billion (world population 6.1 billion)
In India the centre for DNA finger printing and diagnosis (CDFD - centre for DNA finger printing & diagnosis
located at hyderabad
Application of DNA Finger printing
 Paternity tests. The major application of DNA finger printing is in determining family relationships. For
identifying the true (biological) father, DNA samples of Child, Mother and possible fathers are taken and
there DNA finger prints are obtained. The prints of child DNA match to the prints of bilogical parents.
 Identification of the criminal : DNA finger printing has now become useful technique in forensic (crime
detecting science) specially when serious crimes such as murders and rapes are involeved. For
identifying a criminal, the DNA fingerprints of the suspects from blood or hair or semen picked up from
the scene of crime are prepared and compared. The DNA fingerpring of the person matching the one
obtained from sample collected from scene of crime can give a clue to the actual criminal.
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CLONING
Clone is exact carbon copy or copies produced by a single parent (mother or father) clone are produced
by A sexual method and are indentical to their parent genetically and morphologically. Clone is a greek word
which means twig (Klon = twig) deteached shoot. As all the branches of a tree are similar in morphology
and genetical characteristics, in the same way clones are also similar to one another.
Cloning is the process of producing many identical organisms (clone). generally used to produce new plants
with similar characteristics. Microbes produce clones through asexual reproducition. In higher animals,
clones are produced by nuclear transplantion technique in which the nucleus from a sometic cell is
transferred into an unfertilized enucleated egg. The world’s most famous sheep ‘Dolly’ was a clone produced
by this method. Many plant species show vegetative reproduction. In these plants, the clones produced by
a twig (deteached shoot) are similar in their genotype as well as in phenotype (except environmental
variations). Scientists have been much curious to apply this characteristics of plants on animals also to
conserve the descried genotypes of some rare animals by making their clones. In higher animal, showing
sexual reproducition, a zygote is formed after fertilization of the egg by speromatozoan. Zygote differs from
its parents in genotype. It was revealed by the scientists through several experiments that only the egg
or zygote has the potential to produce a whole individual from a single cell. J.B. Gurdon (1969) of Oxford
University applied this fact while performing an experiment on frog. He destroyed the nucleus of an
unfertilized egg of frog by treating with U.V. rays and transferred the nucleus of intestinal epithelial cell
of tadpole into the egg cell. In this experiment a few of the many transplanted eggs could develop into
tadpoles. These developed tadpoles were identical in genotype and phenotype to their parents. This nuclear
transplantation technique by Gurdon is still being used in cloning practice in some modified manner. A brief
intorductory history of cloning is given in table.
In addition to the fact depicted in table attempts are continuously in progress in this field. In December,
2001 (report published in February, 2002) scientists at Univeristy, Taxas, successfully produced the first
cloned domestic pet named as copy cat (C.C.). Further, in Aug. 2005 Woo-Sukhwang of South Korea
produced the clone of an Afganian hound (Domestic dog used for hunting).
An introductory story of cloning
Name of the Brief account of the
Year Result
Scientist experiment (s)

Nuclear transplantation from Tadpoles produced but died

1950 s Briggs & King
embryo to egg in frog. before adulthood.

Nuclear transplantation from

Tadpoles produced but died
1960 s John B. Gurdon cells of skin, liver, kidney into
before adulthood.
the egg in frog.

Nuclear transplantation from Larvae produced but died

1970 s Illmense
embryo to egg in Drosophila before adulthood

Nuclear transplantation from A few mice born but none

1984 Mcgrath & Solter
embryo to egg in mouse lived to adulthood.

Atrificial splitting of an First artificially Twinned

1993 Hall & Stillman embryos developed but embroyos developed but
abnormal abnormal

March Roslin Institute Nuclear transplantation from Megan and Morgan sheep
1995 team, Scotland embryo to egg in sheep born normally

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Roslin institute Nuclear transplantation
Feb, 1997 "Dolly" sheep born normally
team, Scotland udder cell to egg in sheep
Don Wolf and
March Nuclear transplantation from Two monkey "Neti" and
Coworkers,
1997 embryo to egg in monkey "Dolly" sheep born normally
Oregon
Dec., Roslin Institute Nuclear transplanation from "Molly" and Polly sheep
1997 team, Scotland embryo to egg in sheep born normally

University of Nuclear transplantation from "George and Charlie"

1998
Hawai adult cell to egg in cow cows born normally

Well and his Nuclear transplantation from

2000 Many cows born normally
Associated skin cell to egg in cow
Nuclear transplantation from
2001 Kabota skin fibroblast culture to egg Six calves born
in cow

Types of Cloning :
Cloning is an extensive technique, which is divided into following types, on the basis of the experimental
material used :
(1) Gene cloning (2) Microbial cloning (3) Cell cloning
(4) Plant cloning (5) Animal cloning
Animal cloning
Embroyonic cell in animals, lack their totipotency by the time they enter into gastrula stage. So animal
cloning is some what more difficult than plant cloning. On the basis of aims and end products animal
cloning is of two types (i) Reproductive and (ii) Therapeutic.
A clone of the whole animal is prepared in reproductive cloning. The techniques which are used in such
cloning include (1) blastomere separation, (2) nuclear transplantation, (3) Honolulu technique.
This technique can be used to conserve and increase the number of those animals which are threatened
to be extinct in the near future.
In Honolulu technique (1998 - Teruhiko wakayama - cloned mice) nucleus from the donor cell is substituted
into enucleated egg cell (culture medium or chemical both is used in place of electric shock to stimulate
development).
Therapeutic cloning technique may prove to be very useful in the field of medical science, particularly
when there is a need for the transplantation to replace some damaged and diseased organ or tissue by
a healthy organ from a suitable donor. In such condition, if the cells from the patient himself are taken and
cultured to form descried organ. The organs developed in such manner (organ cloning) will be easily
acceptable by the patient, and there will be no possibility of its rejection as often occurs otherwise.
A number of diseases like parkinsonia, alzheimer, diabetes and diseases related with kidneys will possibly
be cured in future by the application of cloning technique. “Dolly” sheep was produced by using nuclear
transfer technique by Dr. Ian Wilmut and his collegues at Roslin Institute of Scotland in 1997. They used
somatic cells from udder (mammary glands) for this clone. One udder cell with its nucleus was selected
because this nucleus carried the mother’s genetic information. Meanwhile, and unfertilized egg cell was

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taken from a different sheep. Its nucleus was sucked out and an enucleated egg cell was obtained. After
then the udder cell nucleus was fused with the enucleated egg cell under electrical stimultation. Now
this egg cell had the mother’s nucleus. At last the fused egg was implanted into the uterus of surrogate
mother, other than the egg donor where it grew into a lamb. Thus the Dolly was born, as a genetically
identical coppy of its mother after the cloning of Dolly, Molly and Polly sheep are cloned by Dr. Ian
Willmut.

Advantages of Cloning
 This technique can be used to improve the breeds of live-stock used in agriculture.
 Cloning is helpful for providing many useful substances and chemicals required for human body, and also
in the cloning of such animals which can be used as a source of organs for transplantation purpose in
medical practices.
 British scientists have been sucessful in obtaining alpha antitrypsin from sheep for curing an incurable
disease emphysema and clotting factor - ix for curing haemophilia. It has been possible by the use of
genetic engineering.
 Cloning is useful to increase the number of individuals of those species which are at the edge of extinction,
thus helpful in conservation of biodiversity.
Disputes Related To Cloning
An Itallian scientist Dr. S. Antenori is trying hard to produce human clone. It has triggred serious discussions
and debates at global level, focussing on the medical and ethical issues.

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At present, the issue of cloning has been a matter of discussion and disputes among the scientists and
the socilogists. Many questions are being with regards to the ethical, moral and social aspects of
cloning. Doubts have also been there about the health and ageing of clones and the misuse of cloning.
POLYMERASE CHAIN REACTION TECHNOLOGY (PCR-TECHNOLOGY)
 This technique was invented by Kary mullis (1983).
 In 1993 Karry Mullis got nobel prize for PCR (for chemistry)
 PCR is a method for amplifying a specific region of DNA molecule without the requirement for time
consuming clonning procedures.
 PCR reaction takes place in Eppendrof tube.
 Using PCR technique very low content of DNA available from sample of blood of semen or any other tissue
or hair cell can be amplified many times and analysed. In this technique Taq-Polymerase is used. Taq
polymerare enzyme is used in PCR which is a special type of DNA polymerare enzyme which is resistant
to high temperature.
 Taq Polymerase is isolated from Thermus aquaticus bacterium.
 Some other examples of polymerase which are used in PCR are –
Pflu Polymerase - Isolated from Pyrococus furiosus bacterium.
Vent Polymerase - Isolated from Thermococcus litoralis bacterium.
3 Main steps in PCR :
(1) Denaturation (2) Annealing (3) Extension
3' 5'

Denaturation (94ºC)

5' 3'

3' 5'
Primer - 1
3' 5'
Annealing (54ºC) 5' 3' Primer - 2
5' 3'

3' 5'

5' 3' 5'

Extension (72ºC)
3' 3' 5'

(Amplified ~ 1 billion
times) 5' 5'
 Denaturation : In this step a double stranded DNA molecule is placed at 94ºC. So double standed DNA
becomes single stranded & each single stranded DNA functions as a template.
 Annealing : In this step two primer DNA are attached at 3’ end of single stranded DNA.
 Extension : In this process taq polymerase enzyme synthesize DNA strain over template. PCR is automatic
process because taq. polymerase enzyme is heat resistant.
 GENOMICS
Genomics is the study of genomes and genes based on DNA seqencing. Genome is the total gene
complement of a haploid set of chromosomes and inherited as a unit from one parent through the gamete.
A haploid (such as prokaryotic) cell contains a single genome, a diploid (such as a cell of higher plant or
animal) has two genomes, one peternal and other maternal. Additional DNA is also present in mitochondria
which is inherited from one’s mother.
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HUMAN GENOME PROJECT
Genetic make-up organism or an individual lies in the DNA sequences. If two individuals differ, then their
DNA sequences should also be different, at least at some places. These assumptions led to the quest of
finding out of the complete DNA sequence of human genome. With the establishment of genetic engineering
techniques where it was possible to isolate and clone any piece of DNA and availability of simple and fast
techniques for determining DNA sequences, a very ambitious project of sequencing human genome was
launched in the year 1990.
Human Genome Project (HGP)
It was a mega project. You can imagine the magnitude and the requriements for the project if we simple
define the aims of the project as follows :
Human genome is said to have approximately 3 × 109 bp, and if the cost of sequencing requred is US $
3 per bp (the estimated cost in the begining), the total estimated cost of the project would be aproximately
9 billion US dollars. Further, if the obtained sequences were to be stored in typed form in books, and if
each page of the book contained 1000 letters and each book contained 1000 pages, then 3300 such such
books would be requred to store the information of DNA sequence from a single human cell. HGP was
closely associated with the rapid development of a new area in biology called as Bioniformatics.
Goals of HGP
Some of the important goals of HGP are as follows :
 Identify all the genes in human DNA.
 Determine the sequences of the 3 billion chemical base pairs that make up human DNA.
 Store this information in database.
 Improve tools for data analysis.
 Transfer related technologies to other sectors. such as industries.
 Address the ethical, legal, and social issues (ELSI) that may arise from the project.
The project was completed in 2003. Knowledge about the effect of DNA variations among individuals can
lead to revolutionary new ways to diagnose, treat and someday prevent the thousands of disorders that
affect human beings. Besides providing clues to understanding human biology, learning about non-human
organisms, DNA sequences can lead to an understanding of their natural capabilities that can be applied
toward solving challenges in health care, agriculture, energy production, environmental remediation. Many
non-human model organisms, such as bacteria, yeast, Caenorhabditis elegans (a freeliving non-pathogenic
nematode), Drosophila (the fruit fly), plants (rice and Arabidopsis), etc., have also been sequenced.
Methodologies : The methods involved two major approaches
 Expressed Sequence Tags : Identifying all the genes by expressed as RNA.
 Sequence Annotation : The blind approach of simply sequencing the whole set of genome that contained
all the coding and non-coding sequence, and later assigning different regions in the sequence, and later
assigining different regions in the sequence with functions. For sequencing, the total DNA from a cell is
isolated and converted into random fragments of relatively smaller sizes (recall DNA is a very long polymer,
and there are technical limitations in sequencing very long pieces of DNA) and cloned in suitable host using
specialised vetors. The cloning resulted into amplification of each piece of DNA fragment so, that is
subsequently could be sequenced with ease. The commonly used hosts were bacteria and yeast, and the
vactors were called as BAC (bacterial artifical chromosomes), and YAC (yeast artificial chromosomes).

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The fragments were sequenced using automated DNA sequencers that worked on the principle of a method
developed by Frederick Sanger. (Remember, Sanger is also credited for developing method for determination
of amino acid sequences in proteins). These sequences were then arranged based on some overlapping
regions present in them. These sequences were then arranged based on some overlapping regions
present in them. This reqired generation of overlapping fragments for sequencing. Alignment of tese sequences
was humanly not possible. Therefore, specialised computer based programmes were developed. These
sequences were subsequently annotated and were assigned to each chromosome. The sequence of
chromosome 1 was completed only in May 2006 (this was the last of the 24 human chromosomes -22
autosomes and X and Y to be sequenced). Another challenging task was assigning the genetic and
physical maps on the genome. This was generated using information on polymorphism of restriction
endonuclease recognition sites, and some repetitive DNA sequences known as microstellites.
Silent Features Of Human Genome :
Some of the salient observations drawn from human genome project are as follows :
 The human genome contains 3164.7 million nucleotide bases.
 The average gene consists of 3000 bases, but sizes vary greatly, with the largest known human gene being
dystrophin at 2.4 million bases.
 The total number of genes is estimated at 30,000 much lower than previous estimates of 80,000 to 1,40,000
genes. Almost all (99.9 percent) nucleotide bases are exactly the same in all people.
 The functions are unknown for over 50 percent of discovered genes.
 Less than 2 per cent of the genome codes for proteins.
 Repeated sequences make up very large protion of the human genome.
 Repetitive sequences are stretches of DNA sequences that are repeated many times, sometimes hundred
to thousand times. They are thought to have no direct coding functions, but they shed light on chromosome
structure, dynamics and evolution.
 Chromosome 1 has most genes (2968). And the Y has the fewest (231).
 Scientists have identified about 1.4 million locations where single base DNA differences (SNPs - single
nucleotide polymorphism, pronounced as ‘snips’) occur in humans, this information promises to
revolutionse the process of finding chromosmal locations for disease-associated sequences and tracing
human history.
Base pair Gene No.
Bateriophage 10,000 - - - - / - - -
Lily 106 Billion B.P. - - - - -
E.coli 4.7 million B.P. 4000
S. cerviceae 12 Million B.P. 6000
D. Melangaster 180 Million B.P. 13,000
Caenorhabdits elegans 97 Million B.P. 18,000
Human 3 Billion B.P. 30,000
 First prokaryote in which complete genome was sequenced is Haemophilus influenzae.
 First Eukaryote in which complete genome was sequenced is Saccharomyces cerviceae (Yeast).
 First plant in which complete genome was sequenced is Arabidopsis thaliana (Small mustard plant).

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 First animal in which complete genome was sequenced is Caenorhabditis elegans (Nematode).
 -globin and insulin gene are less than 10 kilo base pair T.D.F. gene is the smallest gene (14 base pair)
and Duchenne muscular Dystrophy gene is made up of 2400 kilo base pair. (Longest gene)
GENE LIBRARY (Genomic or c-DNA Library) and Gene Banks
A gene library is a collection of many of the desired genes of DNA fragments maintained in clones of
bacterial or some other cells. It is prepared by the following method.
DNA fragments containing one or few desired genes are obtained with the help of specific restriction
endonucleases. Each fragment is joined to a suitable vehicle DNA to form recombinant DNAs of different
nature. These are then intorduced into host (bacterial, yeast plant or animal) cells. The cells containing
recombinant DNAs are allowed to multiply in cultures. This will produce clones of cells where the daughter
cells carry the same genes which are identical to those of parent cells. A collection of clones with
recombinant DNA containing desired genes is a gene library. Gene libraries are maintained through special
techniques.
A gene bank is a store house of clones of known DNA fragments, genes, gene maps, speeds, spores,
frozen sperms or eggs or embryos. These are stored for possible use in genetic engineering and breeding
experiments where spcies have become extinct. The need of gene banks is being increasingly felt as the
rate of extinction is increasing day by day. The human genome project is the most remarkable contribution
in this field.
HUMAN GENETICS
The study (analysis) of genetic characters and aspects like genetic improvements among humans are
included in Human Genetics. This is also knwon as eugenics (=well born).
Eugenics is a term derived from Greek Language Eugenes meaning “Well born”. First of all, Sir Francis
Galton, 1883 proposed the idea of improvement in human species through change in hereditary characters
in a scientific manner and named it Eugenics. Sir Francis Galton is known as “father of Eugenics”. To find
out various facts, scientists have to perform a number of experiments, but it is not possible to do so in
humans. The following problems are faced in studying human genetics :
 It is not possible to perform various experiments on humans in the laboratory.
 Due to greater life span of humans, a lot of time is required to study their genetic characteristics.
 Rate of reproduction is slow in humans.
 Due to controlled hybridization and long lived life among humans, it is not possible to study many
generations in easy way.
Despite of above mentioned problems humans are considered suitable for genetical experiments due
to
 Longer life span of humans, the abnormalities that require relatively long period to express themselves
can be easily studied.
 By pedigree study and analysis of families, many genetic characters of man can be traced out.
 To study many diseases (like haemophilia, colour blindness etc.) and intelligence quatient (I.Q.)
humans are more preferable than any other organisms.
Devices Used In Human Genetical Studies :
The study and analysis of human genetics is performed by many methods like pedigree analysis,
statistical analysis population genetics and human karyotyping. Of these the important ones, that is
pedigree analysis and human karyotype is bing described here.

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 Pedigree Analysis
Study of ancestoral history of man transmission of genetic characters from one generation to next, is
pedigree analysis. Dwarfism, albinism, colour blindness, haemophilia etc. are genetically transmitted
characters. To study and analyse them a pedigree of genetic facts/data and following symbols are used.
Symbol used in Pedigree :

 – Normal Male

 – Narmal Female

 – Mating (Marriage)

Usual practice is to place the males first, on the left if one parent is not shown in a pedigree, it indicates
that this individual was phenotypically normal.

 – The siblings are indicated in chronological order of birth

 – Sex unspecified

 Twin if monozygotic –

 If dizygotic –

 Affected male and female individual

 Heterozygous for autosomal recessive

 Carrier female of sex linked recessive character of disease

 Propositus ; Prooposita – The first patient to be investigated in a family study, to whom

(Probond) (Probond) all relationship are referred.

 – Death of individual

 – Abortion or still birth (sex unspecified)

 – Consanguineous (close relative) marriage

 – Parent with male child affected with disease

 5 – Five unaffected offsprings

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Pedigree analysis provides valuable information regarding genetical make up of human beings. If any
genetic disease is occuring in a family, then pedigree analysis provides guidance to parents about their
future progenies example polydactyly in humans.
 Human Karyotype
Humans have 23 pairs (46) chromosomes. In this method, the chromosomes (autosomes and sex
chromosomes) are arranged according to their size and structure. Based on the position of centromere and
relative length of both arms of chromosome, three types of chromosomes are found in human – metacentric,
submetacentric and acrocentric. Karyotype helps to know the relative structures (morphology) of
chromosomes. Besides, it helps in chromosomal identificaiton. It is also used in studying chromosomal
abnormalities (Down syndrome).
 Types of Eugenics :
For improvement of hereditary characters in future generations there should be an increase in number of
best individuals and reduction in number of defective individuals. Presently, eugenics is studied under
following types :
 Positive Eugencis
Positive eugencis the main approach is to increase the number of progenies having best hereditary characters.
This is achieved through genetic counselling, selective mating (planned marriages), arranging marriages at
right age, freedom from social bonds etc.
Preservation of ovum of superior quality, artifical insemination and preserving the best quality sperms with
aid of modern techniques are also included in positive eugencis.
Euphenics and gene engineering techniques are also being used to establish the best quality of hereditary
traits in human society.
 Negative Eugenics
negative eugencis the carriers of undesirable inferior traits (or characters or genes) are not allowed to
produce their progenies by various checks such as marriage control, segregation, birth control, sterillzation
and control of consanguineous marriages.
Example : Given pedigree shows inheritance of albinism which is an autosumal recessive disorder. If 4th
individul is homozygous normal then find out the carrier individual.
1 2
– Normal male
– Normal Female
4
3 – Albino male
– Albino Female
5 6
Ans : 1, 2, 5, 6 (Aa)

POPULATION GENETICS
Study of gene frequency in a population is called population genetics.
 Gene pool : A gene pool is the sum total of genes in reproductive gametes of a population.
 Gene flow : Migration of gene from one population to another population by cross fertilization.

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 Genetic load : The existance within the population of disadvantageous allele in heterozygous genotype
is known as genetic load
 Gene frequency – Gene frequency is defined as proportion of different alleles of a gene in a population.
Ex. In a population of 200 individuals of MN blood group 100 MM, 40 MN, 60 NN find out the frequency
of M & N.
MM – 100
MN – 40
NN – 60
Total M gene (p) – 100 × 2 + 40 = 240
Total N gene (q) – 60 × 2 + 40 = 160
Total gene = 400
M 240 160
Frequency of M gene (p) P = P = = 0.6 , Frequency of N (q) = = 0.4
MN 400 400
pq 1
Hardy Winberg Law :
1908 G.H. Hardy (English methematician) & German Physician. W. Weinberg independently discovered that
an equilibrium is established between frequencies of allele in random mating population and these gene
frequency remain constant from generation to generation.
This law applicable when factors like mutation, selection & migration, are absent.
Hardy Winberg theoram or Hardy weinbergh law :
P2 + 2pq + q2 = 1 (A  p, a  q, p + q = 1)
AA Aa aa
Equation frequency of A  P so the Frequency of Homozygous dominant will be AA – P2
Equation frequency of a  q so the frequency of homozygous recessive – q2
So, the frequency of Aa = 2pq
The frequency of different genotype produced due to random mating will depend upon the gene frequency
and equilibrium is stablished after one single segeration of random mating.
Factors affecting gene frequency :
(1) Mutation (2) Selection (3) Migration (4) Random drift
 Mutation : Mutation lead to introduction of new gene leading to genetic difference. Change in gene
frequency due to mutation, will depend upon mutation rate.
 Selection (Natural Selection) : By the natural selection there is increase in frequency that are more
fit to nature.
Selections are associated with superior fitness will increase in frequency in the poulation.
 Migration : Migration recessive or dominant allel can be introduced into a population from a nearby
population.
This causes difference in frequency between the two population.
 Random drift or Genetic Drift : Sudden change in gene frequency due to sampling error is called
Genetic Drift.
Change in gene frequency due to random process (gamete fusion) is called Random drift
pq
=
N

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 = sampling error
p & q – initial gene frequency –
N = Total number of genes which are sampled
popultion  sampling error 
population  sampling error 
Q. In a random population frequency of recessive phenotype is 0.09. What is the frequency of heterozygous
genotype ?
Sol. q2 = 0.09;
q = 0.3
p = 1 – q
p = 1 – 0.3  0.7
Frequency of heterozygote = 2pq = 2 × 0.7 × 0.3 = 0.42 = 42%
AFLP (Amplified Fragment Length Polymorphism)
The procedure was described by scientist Zabeau & Vos (1993). In this procedure DNA is cut by restriction
enzyme then these restircted fregment are amplified by P.C.R. and band pattern of these restricted
fragments is visualised after gel electrophoresis.
Procedure (3 steps)
 Digestion of total cellular DNA with one or more restriction enzymes.
 Selective amplication of some of these fragments by P.C.R. primer.
 Electrophoretic separation on gel matrix followed by visualisation of band pattern of these restricted
fragment.
R.A.P.D. (Random Amplification of Polymorphic DNA)
This is a type of P.C.R.in which random (unknown) DNA fragments are amplified.
Bioinformatics :
Definition : Bioinformatics is application of computer technology to the management of bilogical information.
Computer techniques are used togather, store, analyze and integrate biological and genetic information
which can be applied to gene based drug discovery and development.
Drug design Based o Bioinformatics
Bioinformatics is a new approach for drug designing. In this procedure all the knowledge about the disease
is collected, analysed and find out a ‘target molecules’ that cause the disease. Structure of these molecules
is analysed by X-ray N.M.R. technique, then drugs are developed which can bind and block the activity of
these molecules.
Biological Data Base :
Bilogical data base is collection of genomic, proteomic and metabolic data assosiate with computer
software, which include nucleotide sequence of gene or amino acid sequence of protein, Information about
structure, function, location of genes and protein.
Importance :
 Easy asses to the information.
 A method for extracting only required information.

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