Biotechnology

1 Biotechnology Aftab Badshah
BASIC BIOTECHNOLOGY
Definitions
Biotechnology is the use of living systems and organisms to develop or make useful products,
any technological application that uses biological systems, living organisms or derivatives thereof, to make or modify
products or processesforspecific use" Depending on the tools and applications, it often overlaps with the (related) fields
of bioengineering and biomedical engineering.
For thousands of years, humankind has used biotechnology in agriculture, food production, and medicine. The term
itself is largely believed to have been coined in 1919 by Hungarian engineer Károly Ereky. In the late 20th and early
21st century, biotechnology has expanded to include new and diverse sciences such as genomics, recombinant
gene technologies, applied immunology, and development of pharmaceutical therapies and diagnostic tests.
Biotechnology is the integration of natural science and organisms, cells, parts thereof, and molecular analogues for
products and services.
Background
Brewing was an early application of biotechnology.
Agriculture has been theorized to have become the dominant way of producing food since the Neolithic Revolution.
Through early biotechnology, the earliest farmers selected and bred the best suited crops, having the highest yields, to
produce enough food to support a growing population. As crops and fields became increasingly large and difficult to
maintain, it was discovered that specific organisms and their by-products could effectively fertilize, restore nitrogen,
and control pests. Throughout the history of agriculture, farmers have inadvertently altered the genetics of their crops
through introducing them to new environments and breeding them with other plants — one of the first forms of
biotechnology.
These processes also were included in early fermentation of beer. These processes were introduced in
early Mesopotamia, Egypt, China and India, and still use the same basic biological methods. In brewing, malted grains
(containing enzymes) convert starch from grains into sugar and then adding specific yeasts to produce beer. In this
process, carbohydrates in the grains were broken down into alcohols such as ethanol. Later other cultures produced the
process of lactic acid fermentation which allowed the fermentation and preservation of other forms of food, such as soy
sauce. Fermentation was also used in this time period to produce leavened bread. Although the process of fermentation
was not fully understood until Louis Pasteur's work in 1857, it is still the first use of biotechnology to convert a food
source into another form.
For thousands of years, humans have used selective breeding to improve production of crops and livestock to use them
for food. In selective breeding, organisms with desirable characteristics are mated to produce offspring with the same
characteristics. For example, this technique was used with corn to produce the largest and sweetest crops.
Biotechnology has also led to the development of antibiotics. In 1928, Alexander Fleming discovered the
mold Penicillium. His work led to the purification of the antibiotic compound formed by the mold by Howard Florey,
Ernst Boris Chain and Norman Heatley - to form what we today know as penicillin. In 1940, penicillin became available
for medicinal use to treat bacterial infections in humans.
The field of modern biotechnology is generally thought of as having been born in 1971 when Paul Berg's (Stanford)
experiments in gene splicing had early success. Herbert W. Boyer and Stanley N. Cohen significantly advanced the new
technology in 1972 by transferring genetic material into a bacterium, such that the imported material would be
reproduced. The commercial viability of a biotechnology industry was significantly expanded on June 16, 1980, when
the United States Supreme Court ruled that a genetically modified microorganism could be patented in the case
of Diamond v. Chakrabarty. Indian-born Ananda Chakrabarty, working for General Electric, had modified a bacterium
(of the Pseudomonas genus) capable of breaking down crude oil, which he proposed to use in treating oil spills.
A series of derived terms have been coined to identify several branches of biotechnology; for example:
Bioinformatics is an interdisciplinary field which addresses biological problems using computational techniques, and
makes the rapid organization as well as analysis of biological data possible. The field may also be referred to
as computational biology, and can be defined as, "conceptualizing biology in terms of molecules and then applying
informatics techniques to understand and organize the information associated with these molecules, on a large scale.”
Bioinformatics plays a key role in various areas,such as functional genomics, structural genomics, and proteomics, and
forms a key component in the biotechnology and pharmaceutical sector.
Blue biotechnology is a term that has been used to describe the marine and aquatic applications of biotechnology, but
its use is relatively rare.

Green biotechnology is biotechnology applied to agricultural processes. An example would be the selection and
domestication of plants via micro-propagation. Another example is the designing of transgenic plants to grow under
specific environments in the presence (or absence) of chemicals. One hope is that green biotechnology might produce
more environmentally friendly solutions than traditional industrial agriculture. An example of this is the engineering of
a plant to express a pesticide, thereby ending the need of external application of pesticides.
Red biotechnology is applied to medical processes. Some examples are the designing of organisms to
produce antibiotics, and the engineering of genetic cures through genetic manipulation.
White biotechnology, also known as industrial biotechnology, is biotechnology applied to industrial processes. An
example is the designing of an organism to produce a useful chemical. Another example is the using of enzymes as
industrial catalysts to either produce valuable chemicals or destroy hazardous/polluting chemicals. White biotechnology
tends to consume less in resources than traditional processes used to produce industrial goods.
Biotechnology and its Various Stages of Development
There are various stages in the development of biotechnology to meet the various needs of humans. Its development
was basically based on observations, and applications of these observations to practical scenarios. The complexity of
biotechnology is augmented due to evolution of new technologies with time, as these are based on the employment of
improved technological advancements along with better understanding of various principles of life-science. If, we
systemically study the developments of biotechnology up to its current stage,it can be divided into three different stages
or categories: (1) Ancient Biotechnology, (2) Classical Biotechnology, and (3) Modern Biotechnology.
Ancient Biotechnology (Pre-1800)
Most of the developments in the ancient period i.e., before the year 1800, can be termed as ‘discoveries’ or
‘developments’. If we study all these developments, we can conclude that all these inventions were based on common
observations about nature, which could be put to test for the betterment of human life at that point in time.
Food, clothes, and shelter are the most important basic needs of human beings irrespective of whether they lived in the
ancient period or the modern period. The only factor that has changed is their types and origins. Early man used to eat
raw meat. However,during harsh weather,there was a paucity of food which led to the domestication of food products,
which is named as ‘agriculture’. In ancient times, humans explored the possibilities of making food available by growing
it near their shelters, so that the basic need for food could be met easily. They brought seeds of plants (mostly grains)
and sowed them near to their shelters. Domestication of wild animals was the beginning of observation, implications,
and applications of animal breeding. Certainly, we can say that this was the initial period of evolution of farming, which
led to another needslike the development of methods for food preservation and storage.They used cold cavestopreserve
food for long-term storage. It also made the way for the evolution of pots to store food products, in the form of leather
bags, clay jars, etc.
Certainly, cheese can be considered as one of the first direct products (or by-product) of biotechnology, because it was
prepared by adding rennet (an enzyme found in the stomach of calves) to sour milk, which is possible only by exposing
milk to microbes. Yeast is one of the oldest microbes that have been exploited by humans for their benefit. Yeast has
been widely used to make bread, vinegar production, and other fermentation products, which include production of
alcoholic beverages like whiskey, wine, beer, etc.
Classical Biotechnology
The second phase of evolution and development of biotechnology can be called ‘Classical Biotechnology’. This phase
existed from 1800 to almost the middle of the twentieth century. During this period various observations started pouring
in, with scientific evidences. They were all very helpful toward solving the puzzles of biotechnology. Each and every
contribution from different individuals helped to solve the puzzle and pave the path for new discoveries.
The basics for the transfer of genetic information are the core of biotechnology. This was,for the first time, interpreted
in plants, i.e., Pisum sativum, commonly known as Pea plant. These observations were decoded by Gregor John Mendel
(1822-1884), anAustrian Augustinian Monk. Mendelat that time presented“Laws of Inheritance” to the NaturalScience
Society in Brunn, Austria. Mendel proposed that invisible internal units of information account for observable traits,
and that these ‘factors’ -later called as genes, which are passed from one generation to the next.
Robert Brown had discovered nucleus in cells, while in 1868, Fredrich Miescher, a Swiss biologist reported nuclein, a
compound that consisted of nucleic acid that he extracted from pus cells i.e., white blood cells (WBC). These two
discoveries became the basis of modern molecular biology, for the discovery of DNA as a genetic material, and the role

of DNA in transfer of genetic information. 1n 1881, Robert Koch, a German physician described the bacterial colonies
growing on potato slices. In 1888, Waldeyer-Hartz, a German scientist coined the term ‘Chromosome’, which is
considered as an organized structure of DNA and protein present in cells or a single piece of coiled DNA containing
many genes, regulatory elements, and other nucleotide sequences. Other important discoveries during this period were
vaccination against small pox and rabies developed by Edward Jenner a British Physician and Louis Pasteur a French
Biologist.
By this time the development and growth of biological sciences seemed to be reaching to the exponential phase. The
principle of genetics in inheritance was redefined by T H Morgan, who has shown inheritance and the role of
chromosomes in inheritance by using fruit flies, i.e., Drosophila melanogaster. This landmark work of T H Morgan was
named, ‘The theory of the Gene’ in 1926.
Almost at the same time, in Britain, Alexander Fleming a physician discovered antibiotics, when he observed that one
microorganism can be used to kill another microorganism, a true representation of the ‘divide and rule’ policy of
humans. Fleming noted that all bacteria (Staphylococci) died when a mold was growing in a petri-dish. Later he
discovered ‘penicillin’ the antibacterial toxin from the mold Penicillium notatum, which could be used against many
infectious diseases.
Modern Biotechnology
In 1953, JD Watson and FHC Crick for the first time cleared the mysteries around the DNA as a genetic material, by
giving a structural model of DNA,popularly known as,‘Double Helix Model of DNA’. This model was able to explain
various phenomena relatedto DNAreplication, and its role in inheritance. Later,Jacob and Monad had given the concept
of Operon in 1961, while Kohler and Milestein in 1975, came up with the concept of cytoplasmic hybridization and
produced the first ever monoclonal antibodies, which has revolutionized the diagnostics.
Dr. Hargobind Khorana was able to synthesize the DNA in test tube, while Karl Mullis added value to Khorana's
discovery by amplifying DNA in a test tube, thousand times more than the original amount of DNA. Using this
technological advancement, other scientists were able to insert a foreign DNA into another host and were even able to
monitor the transferof a foreign DNAinto the next generation. The advent of HIV / AIDSasa deadly disease has helped
tremendously to improve various tools employed by life-scientist for discoveries and applications in various aspects of
day-to-day life. In the meantime Ian Wilmut an Irish scientist was successfulto clone an adult animal, using sheep as
model, and he named the cloned sheep as ‘Dolly’. Craig Venter,in 2000, was able to sequence the human genome; the
first publically available genome is from JD Watson and Craig Venter, himself. These discoveries have unlimited
implications and applications.
History of Biotechnology
1855: The Escherichia coli bacterium is discovered. It later becomes a major research,development, and production
tool for biotechnology. Pasteur begins working with yeast, eventually proving they are living organisms.
1888: The chromosome is discovered by Waldyer.
1909: Genes are linked with hereditary disorders.
1915: Phages, or bacterial viruses, are discovered.
1919: The word "biotechnology" is first used by a Hungarian agricultural engineer.
1928: Fleming discovers penicillin, the first antibiotic.
1941: The term "genetic engineering" is first used by a Danish microbiologist.
1942: The electron microscope is used to identify and characterize a bacteriophage- a virus that infects bacteria.
1953: Watson and Crick reveal the three-dimensional structure of DNA.
1958: DNA is made in a test tube for the first time.
1967: The first automatic protein sequencer is perfected.
1969: An enzyme is synthesized in vitro for the first time.
1973: Cohen and Boyer perform the first successful recombinant DNA experiment, using bacterial genes.
1978: Recombinant human insulin is produced for the first time. Human growth hormone is synthesized for the first
time.

1980: The U.S. Supreme Court approves the principle of patenting organisms, which allows the Exxon oil company
to patent an oil-eating microorganism. 1980: Smallpox is globally eradicated following 20-year massvaccination effort.
1981: The North Carolina Biotechnology Center is created by the state's GeneralAssembly as the nation's first state-
sponsored initiative to develop biotechnology. The first gene-synthesizing machines are developed. The first
genetically engineered plant is reported. Mice are successfully cloned.
1982: The first biotech drug, human insulin produced in genetically modified bacteria, is approved by FDA.
Genentech and Eli Lilly developed the product. The first recombinant DNA vaccine for livestock is developed.
1983: The Polymerase Chain Reaction (PCR) technique is conceived. PCR, which uses heat and enzymes to make
unlimited copies of genes and gene fragments, later becomes a major tool in biotech research and product development
worldwide. The first genetic transformation of plant cells by TI plasmids is performed. The first artificial chromosome
is synthesized. The first genetic markers for specific inherited diseases are found. Efficient methods are developed to
synthesize double-stranded DNA from first-strand cDNA involving minimal loss of sequence information.
1986: The first recombinant vaccine for humans, a vaccine for hepatitis B, is approved. Interferon becomes the first
anticancer drug produced through biotech.
1988: Congress funds the Human Genome Project, a massive effort to map and sequence the human genetic code as
well as the genomes of other species. The first pest-resistant corn, Bt corn, is produced.
1990: The first federally approved gene therapy treatment is performed successfully on a 4-year old girl suffering
from an immune disorder.
1993: Chiron's Betaseron is approved as the first treatment for multiple sclerosis in 20 years. The FDA declares that
genetically engineered foods are "not inherently dangerous" and do not require special regulation.
1994: The first breast cancer gene is discovered.
1996: A gene associated with Parkinson’s disease is discovered. The first genetically engineered crop is
commercialized.
1997: Scottish scientists report cloning a sheep, using DNA from adult sheep cells. A group of Oregon researchers
claims to have cloned two Rhesus monkeys. A new DNA technique combines PCR, DNA chips, and a computer
program, providing a new tool in the search for disease-causing genes.
1998: A rough draft of the human genome map is produced, showing the locations of more than 30,000 genes .Cloned
vain RT with fully active polymerase and minimized RNAse H activity is engineered.
2001: The sequence of the human genome is published in Science and Nature, making it possible for researchers all
over the world to begin developing treatments.
2003: The Human Genome Project completes sequencing of the human genome. China grants the world’s first
regulatory approval of a gene therapy product, Gendicine (Shenzhen SiBiono GenTech), which delivers the p53 gene
as a therapy for squamous cell head and neck cancer.
2004: UN Food and Agriculture Organization endorses biotech crops, stating biotechnology is a complementary tool
to traditional farming methods that can help poor farmers and consumers in developing nations.
2006: The National Institutes of Health begins a 10-year,10,000-patient study using a genetic testthat predicts breast-
cancerrecurrence andguides treatment. Patients whose cancer is deemed unlikely to recur will be sparedchemotherapy.
FDA approves the recombinant vaccine Gardasil®, the first vaccine developed against human papillomavirus (HPV),
an infection implicated in cervical and throat cancers, and the first preventative cancer vaccine. The genetic test,
Oncotype DXTM was developed by the biotech company Genomic Health and is already commercially available.
2010: Researchers at the J. Craig Venter Institute create the first synthetic cell.
The scope and importance of Biotechnology:
Biotechnology may be as old as human civilization but modern biotechnology is less than three decades old. Traditional
Biotechnology that led to the development of processes for producing products like yogurt, Vinegar, alcohol and cheese
was entirely empirical and bereft of any understanding of the mechanisms that led to the product. There was no
possibility of a deliberate design to produce a desired new product.
In modern biotechnology, we use the in- depth understanding we have gained in the last five decades. The mechanisms
that underlie the variety of functions performed by living organisms, to produce a desired new or old product. In the
case of an established product, the new biotechnological process is cheaper and better in many respects than the earlier

processes.
Modern biotechnology hasbeen, infact, an historical imperative. Its emergence on the world scene waspredicted atleast
four decades ago. The term, genetic engineering, was coined independently in 1973 by the author of an article in The
Guardian in the UK, and in a syndicated article by the present author in India.
Today's biotechnology has being characterized by the use of a different set of technologies.
Scope ofbiotechnology
Genetic engineering: Genetic Engineering of microbes, plants and animals (including marine animals). Genetic
engineering implies conferring new capabilities on an organism by transferring into an organism the
appropriate DNA (De oxyribo Nucleic Acid, the genetic material) of another having these capabilities does this. Then
ensures that these capabilities are converted into abilities. Thus the common yeast, Sacchromyces cerevisciae cannot
make the protein, human insulin, but we can make it to do so by introducing in it the gene for human insulin (that is,
the appropriate DNA fragment coding for this protein). After integrating the insulin gene in yeast DNA, creates
condition for the insulin to express itself to produce insulin through the normal process of transfer of information
from DNA to protein.
Genetically engineered microbes are today widely used for producing drugs and vaccines in large scale at low costs
that are of great importance (human insulin, erythropoietin, and hepatitis-B vaccine). Genetically engineering plants
are also poised to produce vaccines. A few hundred acres of genetically engineered banana plantation can provide
enough vaccine to immunize 120 million children every year that need to be protected against four common diseases.
One of the future sources of cheap protein-drugs in the coming years, would be genetically engineered animals who
would secrete these drugs in abundance (1-15 mg/ml) in their milk. They will be available at a cost of three or more
times lower than the current cost.
Gene Therapy: This is in a way, genetic engineering of humans, which would allow a person suffering from
a disabling genetic disorder to lead a normal life.
Immuno-technologies: Such as monoclonal antibodies (MABs) for diagnosis and therapy. Antibodies, special sets of
proteins present in humans that enable them to fight incursion of their bodies by harmful chemicals or micro-
organisms. Monoclonal antibodies are single chemical species of antibodies produced in the laboratory by a special
technique. Nobel Prize was awarded for this in the 1980's to Cesar Milstein and Georges Kashler. Mouse MAB's can
be used for the diagnosis of human diseases. As human MABs are difficult to produce in the laboratory, genetically
engineered plants are likely to find wide application in the production of human MABs.
Tissue culture: Tissue culture of both plant and animal cells. These are used for Micro propagation of elite or
exotic materials (Such as orchids), production of useful compounds such as taxol (the widely used anti-cancer drug)
and vanillin, and preparation in the laboratory of "natural" tissues such as arteries for arterial graft or skin for burn
victims. (Modern tissue culture technologies allow the multiplication in the laboratory of cells isolated from plants and
animals. In the case of plants, one can grow in the lab a whole plant from a single cell.)
Stem cell techniques: Which would involve purification and isolation of stem cells from various tissues and
develop into the desired tissue which could then be used, for example, for transplantation. Stem cells can be either
totipotent (have the capability to produce any desired cell type or organ of the body under specific conditions) or they
could be pluripotent (able to develop into severalthough not all cell types or organs). As embryonic stem cells are
more likely totipotent than stem cells from adult tissues, the immediate emphasis in the area of stem cells is going to
be first in the direction of establishing cell lines derived from early human embryos, from which stem cells could be
isolated.
Enzyme engineering and technology: Involves immobilized or stabilized enzymes, new classes of enzymes
(ribozymes) or new enzymatic routes that produce important organic compounds. Enzymes are biological catalysts
(Generally proteins) poised to replace inorganic catalysts, which are used in chemical industry. (Proteins are abundant
biological entities made up of twenty amino acids strung together like pearls in a necklace, by a special type of thread-
a chemical bond called the peptide bond. One protein differs from another in the total number of amino acids and their
sequence in the chain.)
Photosynthetic efficiency: Increasing photosynthetic efficiency for biomass production in the plant with the
same amount of light and other inputs.
NewDNA technologies: These include DNA fingerprinting, sequencing of genomes, development and use of

new molecular markers for plant identification and characterization. Also the development of DNA- based probes for
diagnosis of inherited disorders, antisense technologies that are aimed at blockage of the function of a particular
stretch of DNA and computing using DNA.
Plant-based drugs: Use of modern biological techniques for validation, standardization and manufacture of
indigenous plant-based drug formulations.
Peptide synthesis: Synthesis to make new drugs or other materials of industrial and commercial importance,
such as salmon GnRH analogue (Ovaprin) to induce ovulation in fish.
Rational drug design: Until a decade or so ago, the only way to discover a newdrug wasto synthesize a large number
of compounds hoping that one of them will be effective against a particular disease. In rational drug design, we first
identify the molecular targetwe wish to attack. To do so, it becomes necessarytounderstand the mechanism of causation
of the disease. Once we understand this mechanism and identify the molecular target lead effective computerized
programs to design a molecule, which would hit the target.
Nutraceuticals: That helps recovery after surgery or an episode of a major disease, or helps protect one against
certain medical and health problems. For example, a Swedish company, Probi, has isolated a strain of Lactobacillus
planetarum, which is apparently present in the digestive tract of Europeans and Amercians. The presence of this
organism has been correlated with the ability of the person to recover after major surgery or after chemotherapy of
cancer; this organism also seems to protect people against a vast range of stomach disorders including stomach ulcers,
irritable bowel syndrome and constipation. Probi is, therefore, marketing this organism in various forms, including a
delicious soft drink!
Assisted reproductive technologies: Such as artificial insemination (Using husband's or donor semen), invitro
fertilization, intra cytoplasmic sperm injection and techniques involving egg donation, surrogate motherhood or embryo
transfer.
New cloning technologies: Cloning of genetically engineered animals that would produce useful products.
Organ transplantation: Xenotransplantation that is transplantation into humans of organs from other animals.
It appears that pig may be the most suitable for this biochemically, anatomically and immunologically. The major
problem in xenotransplantation is the hyper-acute immunological rejection of the “foreign organ" which occurs in a
matter of minutes. This problem has been recently overcome by identifying the molecular basis of the hyper-acute
rejection and then genetically engineering a pig to avoid it.
New drug-delivery systems: Such as lipsomes and electrical patches, and the use of circadian rhythms to optimize
the effectiveness of the drug. Thus the drug may depending on the circadian rhythm of the individual will be effective
when taken at noon and midnight, than if taken at 6 AM and 6 PM.
Bioremediadtion: For example of effluents or waste, using biological systems. A septic tank and an oxidation
pond are simple examples of such bioremediation. Production of biogas is value-added bioremediation!
Biological warfare: This is defined as the 'employment of biological agentsto produce casualties in man or animals
or damage to plants. While a biological attackcould result in a made-made epidemic of unprecedentedscale, the classical
principles of clinical medicine and epidemiology would apply. Prompt diagnosis and early interventions could reduce
morbidity and mortality, and mitigate the effects of a biological attack

Biotechnology

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