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An English Medium Co. Ed. Senior Secondary School
Investigatory Project
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Aknowledgement
I am overwhelmed in all humbleness and gratefulness to acknowledge my depth to all
those who have helped me to put these ideas, well above the level of simplicity and into
something concrete.
I would like to express my special thanks of gratitude to my biology teacher, Mr. Sandeep
Kulshesthra as well as our Principal Mrs. Nidhi Bhatia who gave me the golden
opportunity to do this wonderful project on the topic “Applications of Biotechnology”,
which also helped me in doing a lot of research and I came to know about so many new
things. I am really thankful to them.
Any attempt at any level can’t be satisfactorily completed without the support and
guidance of my Parents and Friends who helped me a lot in gathering different
information, collecting data and guiding me from time to time in making this project,
despite of their busy schedules, they gave me different ideas in making this project
unique. I am thankful to them too.
I am making this project not only for marks but to also increase my knowledge...
Thanking you
Subhag Singh
XII Sci. B
Certificate
This is to certify that SUBHAG SINGH of class XII SCI.B of
GYAN DEEP SHIKSHA BHARATI has successfully completed
the investigatory project on the topic “APPLICATIONS OF
BIOTECHNOLOGY” under the guidance of MR. SANDEEP
KULSHESTHRA (H.O.D. Biology) during the session 2015-
16 in the partial fulfilment of Biology Practical
Examination conducted by CENTRAL BOARD OF
SECONDARY EDUCATION (AISSCE).
___________________ ___________________
Mr. Sandeep Kulshesthra External Examiner
(H.O.D Biology) (C.B.S.E)
Introduction
What is Biotechnology?
Biotechnology is the use of living systems and organisms to develop or make products, or
"any technological application that uses biological systems, living organisms or derivatives
thereof, to make or modify
products or processes for
specific use.
At its simplest,
biotechnology is technology
based on biology -
biotechnology harnesses
cellular and bio molecular
processes to develop
technologies and products
that help improve our lives
and the health of our planet.
We have used the biological
processes of
microorganisms for more
than 6,000 years to make
useful food products, such as bread and cheese, and to preserve dairy products.
Modern biotechnology provides breakthrough products and technologies to combat
debilitating and rare diseases, reduce our environmental footprint, feed the hungry,
useless and cleaner energy, and have safer, cleaner and more efficient industrial
manufacturing processes.
Biotech is helping to heal the world by harnessing nature's own toolbox and using our
own genetic makeup to heal and guide lines of research by:
 Reducing rates of infectious disease
 Saving millions of children's lives
 Changing the odds of serious, life-threatening conditions affecting millions around the
world
 Tailoring treatments to individuals to minimize health risks and side effects
 Creating more precise tools for disease detection
 Combating serious illnesses and everyday threats confronting the developing world.
History
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. 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 mould Penicillium. His work led to the purification of the
antibiotic compound formed by the mould 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 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.
Biotechnology in Agriculture
Genetically Modified Crops
Genetically modified crops or “GM crops” or
“biotech crops” are plants used in agriculture,
the DNA of which has been modified with genetic
engineering techniques. In most cases the aim is to
introduce a new trait to the plant which does not
occur naturally in the species. Examples in food
crops include resistance to certain
pests, diseases, stressful environmental
conditions, resistance to chemical treatments,
reduction of spoilage, or improving the nutrient profile of the crop. Examples in non-food
crops include production of pharmaceutical agents, bio fuels, and other industrially useful
goods,as well as for bioremediation.
Plants and crops with GM traits have been tested more than any other crops—with no
credible evidence of harm to humans or animals. In fact, seeds with GM traits have been
tested more than any other crops in the history of agriculture – with no credible evidence
of harm to humans or animals.
Governmental regulatory agencies, scientific organizations and leading health
associations worldwide agree that food grown from GM crops is safe to eat. The World
Health Organization, the American Medical Association, the U.S. National Academy of
Sciences, the British Royal Society, among others that have examined the evidence, all
come to the same conclusion: consuming foods containing ingredients derived from GM
crops is safe to eat and no riskier than consuming the same foods containing ingredients
from crop plants modified by conventional plant improvement techniques.
Genetic modifications have:
1. Made crops more tolerant to abiotic stresses (cold, drought, salt, heat).
2. Reduced reliance on chemical pesticides (pest resistant crops).
3. Helped to reduce post harvest losses & enhanced the nutritional value of the food.
RNA Interference (RNAi)
RNA interference (RNAi) is a method of blocking gene function by inserting short
sequences of ribonucleic acid (RNA) that match part of the target gene’s sequence, thus
no proteins are produced. RNAi has the potential to become a powerful therapeutic
approach toward targeted and personalized medicine. RNAi has provided a way to control
pests and diseases, introduce novel plant traits and increase crop yield. Using RNAi,
scientists have developed novel crops such as nicotine-free tobacco, non-allergenic
peanuts, decaffeinated coffee, and nutrient fortified maize among many others.
Mechanism of RNA interferences as understood is that it comes into play when a double
stranded RNA is introduced either naturally or artificially in a cell. An endo ribonuclease
enzyme cleaves the long dsRNA into small pieces of RNA. The small pieces could be mi
RNA or si RNA depending upon the origin of long dsRNA i.e. endogenous or exogenous
respectively. A double stranded RNA may be generated by either RNA dependent RNA
polymerase or bidirectional
transcription of transposable
elements or physically introduced.
There are several opportunities for
the applications of RNAi in crop
science for its improvement such as
stress tolerance and enhanced
nutritional level.This knockdown
technology may be useful in inducing
early flowering, delayed ripening,
delayed senescence, breaking
dormancy, stress-free plants,
overcoming self-sterility, etc.
RNA interference (RNAi) has recently been demonstrated in plant parasitic nematodes. It
is a potentially powerful investigative tool for the genome-wide identification of gene
function that should help improve our understanding of plant parasitic nematodes. RNAi
should help identify gene and, hence, protein targets for nematode control strategies.
Prospects for novel resistance depend on the plant generating an effective form of
double-stranded RNA in the absence of an endogenous target gene without detriment to
itself. These RNA molecules must then become available to the nematode and be capable
of ingestion via its feeding tube. If these requirements can be met, crop resistance could
be achieved by a plant delivering a dsRNA that targets a nematode gene and induces a
lethal or highly damaging RNAi effect on the parasite.
Bt toxin
A protein that is toxic to chewing insects and is produced by the soil bacterium Bacillus
thuringiensis and has long been used as a biological pesticide. By means of genetic
engineering, the genes for Bt toxin can be isolated from Bacillus thuringiensis and
transferred to plants.
Bacillus thuringiensis (Bt) is a bacteria that produces proteins which are toxic to insects.
But extreme toxicity comes at no surprise. It’s in the same family of bacteria as B.
anthracis, which causes anthrax, and B. cereus, which causes food poisoning.
The Bt toxin dissolve in the high pH insect gut and become active. The toxins then attack
the gut cells of the insect, punching holes in the lining. The Bt spores spills out of the gut
and germinate in the insect causing death within a couple days.
Even though the toxin does not kill the insect immediately, treated plant parts will not be
damaged because the insect stops feeding within hours. Bt spores do not spread to other
insects or cause disease outbreaks on their own.
1. Insect eats Bt crystals and spores.
2. The toxin binds to specific receptors in the
gut and the insects stops eating.
3. The crystals cause the gut wall to break
down, allowing spores and normal gut bacteria
to enter the body.
4. The insect dies as spores and gut bacteria
proliferate in the body.
Bt action is very specific. Different strains of Bt are specific to different receptors in insect
gut wall. Bt toxicity depends on recognizing receptors, damage to the gut by the toxin
occurs upon binding to a receptor. Each insect species possesses different types of
receptors that will match only certain toxin proteins, like a lock to a key.
It is because of this that farmers have to be careful to match the target pest species with
a particular Bt toxin protein which is specific for that insect. This also helps the benifical
insects because they will usually not be harmed by that particular strain of Bt.
Bt Cotton
Bt cotton is a genetically modified organism (GMO) cotton variety, which produces
an insecticide to bollworm. Strains of the bacterium Bacillus thuringiensis produce over
200 different Bt toxins, each harmful to different insects. Most notably, Bt toxins are
insecticidal to the larvae of moths and
butterflies, beetles, cotton bollworms and ghtu flies but are
harmless to other forms of life. The gene coding for Bt toxin
has been inserted into cotton as a transgene, causing it to
produce this natural insecticide in its tissues. In many
regions, the main pests in commercial cotton
are lepidopteran larvae, which are killed by the Bt protein in
thegenetically modified cotton they eat. This eliminates the
need to use large amounts of broad-spectrum insecticides to
kill lepidopteran pests. This spares natural insect predators
in the farm ecology and further contributes to non
insecticide pest management.
Bt cotton is ineffective against many cotton pests such as plant bugs, stink bugs,
and aphids; depending on circumstances it may be desirable to use insecticides in
prevention. A 2006 study done by Cornell researchers, the Center for Chinese Agricultural
Policy and the Chinese Academy of Science on Bt
cotton farming in China found that after seven years
these secondary pests that were normally controlled by
pesticide had increased, necessitating the use of
pesticides at similar levels to non-Bt cotton and causing
less profit for farmers because of the extra expense of
GM seeds.
Mechanism:
Bt cotton was created through the addition of genes
encoding toxin crystals in the Cry group
of endotoxin. When insects attack and eat the cotton
plant the Cry toxins are dissolved due to the high pH level of the insects stomach. The
dissolved and activated Cry molecules bond to cadherin-like proteins on cells comprising
the brush border molecules. The epithelium of the brush border membranes separates
the body cavity from the gut whilst allowing access for nutrients. The Cry toxin molecules
attach themselves to specific locations on the cadherin-like proteins present on the
epithelial cells of the midge and ion channels are formed which allow the flow of
potassium. Regulation of potassium concentration is essential and, if left unchecked,
causes death of cells. Due to the formation of Cry ion channels sufficient regulation of
potassium ions is lost and results in the death of epithelial cells. The death of such cells
creates gaps in the brush border membrane.
Advantages:
Bt cotton has several advantages over non Bt cotton. The important advantages of Bt
cotton are briefly :
 Increases yield of cotton due to effective control of three types of bollworms, viz.
American, Spotted and Pink bollworms.
 Insects belonged to Lepidoptera (Bollworms) are sensitive to crystalline endotoxic
protein produced by Bt gene which in turn protects cotton from bollworms.
 Reduction in pesticide use in the cultivation of Bt cotton in which bollworms are major
pests.
 Reduction in the cost of cultivation and lower farming risks.
 Reduction in environmental pollution by the use of insecticides rarely.
 Bt cotton exhibit genetic resistance or inbuilt resistance which is a permanent type of
resistance and not affected by environmental factors. Thus protects crop from
bollworms.
 Bt cotton is ecofriendly and does not have adverse effect on parasites, predators,
beneficial insecticides and
organisms present in soil.
 It promotes multiplication of
parasites and predators
which help in controlling the
bollworms by feeding on
larvae and eggs of bollworm.
 No health hazards due to
rare use of insecticides.
 Bt cotton are early in
maturing as compared to
non Bt cotton.
Disadvantages:
Bt cotton has some limitations
 High cost of Bt cotton seeds as compared to non Bt cotton seeds.
 Effectiveness up to 120 days, after that the toxin producing efficiency of the Bt gene
drastically reduces.
 Ineffective against sucking pests like jassids, aphids, whitefly etc.
Bt cotton in India:
Bt cotton is supplied in India's Maharashtra state by the agri-biotechnology company,
Mahyco, as the distributor.
The use of Bt cotton in India has grown exponentially since its introduction. Recently India
has become the number one global exporter of cotton and the second largest cotton
producer in the world. India has bred Bt-cotton varieties such as Bikaneri Nerma and
hybrids such as NHH-44, setting up India to benefit now and well into the future.
India’s success has been subject to scrutiny. Monsanto's seeds are expensive and lose
vigour after one generation, prompting the Indian Council of Agricultural Research to
develop a cheaper Bt cotton variety with seeds that could be reused. The cotton
incorporated the cry1Ac gene from the soil bacterium Bacillus thuringiensis (Bt), making
the cotton toxic to bollworms. In parts of India cases of acquired resistance against Bt
cotton have occurred.
The state of Maharashtra banned the sale and distribution of Bt cotton in 2012, to
promote local Indian seeds, which demand less water, fertilizers and pesticide input, but
lifted the ban in 2013.
India approved Bt cotton in 2002; now it accounts for 92% of all Indian cotton. Average
nationwide cotton yields went from 302 kg/ha in the 2002/3 season to a projected 481
kg/ha in 2011/12 — up 59.3% overall. This chart shows the trends in yields, which took
off after Bt was introduced in 2002. The graphs also show that — and here comes ugly
fact— in the last 4 years, as Bt has risen from 67% to 92% of India’s cotton, yields have
dropped steadily.
Biotechnology in
Medicine
Genetically Engineered Insulin (Humulin)
Insulin is a peptide hormone produced by beta
cells in the pancreas of various organisms
including human beings. It regulates
the metabolism of carbohydrates and fats by
promoting the absorption of glucose from the
blood to skeletal muscles and fat tissue and by
causing fat to be stored rather than used for
energy. Insulin also inhibits the production of
glucose by the liver.
Except in the presence of the metabolic disorder diabetes mellitus and metabolic
syndrome, insulin is provided within the body in a constant proportion to remove excess
glucose from the blood, which otherwise would be toxic. When blood glucose levels fall
below a certain level, the body begins to use stored glucose as an energy source
through glycogenolysis, which breaks down the glycogen stored in the liver and muscles
into glucose, which can then be utilized as an energy source. As a central metabolic
control mechanism, its status is also used as a control signal to other body systems (such
as amino acid uptake by body cells). In addition, it has several other anabolic effects
throughout the body. When control of insulin levels fails, diabetes mellitus can result.
Structure:
Insulin is composed of two different
types of peptide chains. Chain A has 21
amino acids and Chain B has 30 amino
acids. Both chains contain alpha
helices but no beta strands. There are 3
conserved disulfide bridges which help
keep the two chains together. Insulin
can also form dimers in solution due to
the hydrogen bonding between the B
chains. The dimers can further interact
to form hexamers due to interaction between hydrophobic surfaces. This scene
highlights the hydrophobic and polar parts of an insulin monomer at a pH of 7.
A number of insulin variants have been made to favor either the monomeric or hexameric
form. Deletion of the five C terminal residues of the B chain creates a monomer only
form. This portion of the B chain is involved in hydrogen bonds between the B chain of
one monomer and the A and B chain of another monomer.
Need of Genetically Engineered Insulin:
The original form of the wonder cure for diabetes, these were once the only type of
insulin available, but are now rarely used. Animal insulin was originally made from
ground-up animal pancreas tissue, and then later was extracted from healthy animals
(slaughtered pigs & cows).
The metabolism of cows
and pigs was close
enough to human
metabolism that their
animal insulin also
worked well in human
bodies. Beef insulin has 3
differences from human; pork insulin has 1 difference from human. The use of a mixture
of beef and pork insulin was also possible. It has been shown that human insulin is less
immunogenic than animal insulin. Porcine insulin is most similar to human insulin. The
primary amino acid sequences of bovine and porcine insulin differ from that of human
insulin by three and one amino acid, respectively. This greater dissimilarity between
human and bovine insulin has been postulated to be the explanation for the greater
antigenicity of bovine insulin as compared with porcine insulin
One of the problems with animal insulin was antibody issues. The body identifies them
and tries to reject them. Pork insulin differs by 1 amino acid and beef insulin by 3 amino
acids, so the body's immune system can sometimes
recognize them as foreign. Immunological complications
of insulin therapy have been evident since animal insulin
became available for the treatment of diabetes mellitus in
1922. In insulin-allergic patients treated with conventional
insulin preparations, the insulin-specific IgE values are
often 10- to 20-fold higher than in patients without
allergy. It has been shown that human insulin is less
immunogenic than animal insulin. Porcine insulin is most
similar to human insulin. Cross-reactivity between human insulin and insulin of animal
origin has been reported. A major problem is the cross-reactivity that occurs between
anti-insulin antibodies and the various animal and human insulin preparations in patients
presenting with allergy to animal insulin.
The usage of animal insulin has so greatly declined in modern times that they have
largely been withdrawn from the market. Newly diagnosed diabetics are typically given
synthesized or Genetically Engineered human insulin.
What is “Proinsulin”?
Proinsulin is the prohormone precursor to insulin made in the beta cells of the islets of
Langerhans, specialized regions of the pancreas. Proinsulin is synthesized on membrane
associated ribosomes found on the
rough endoplasmic reticulum, where it
is folded and its disulfide bonds are
oxidized. It is then transported to
the Golgi apparatus where it is
packaged into secretory vesicles, and
where it is processed by a series of
proteases to form mature insulin.
Mature insulin has 35 fewer amino
acids; 4 are removed altogether, and
the remaining 31 form the C-peptide.
The C-peptide is abstracted from the
center of the proinsulin sequence; the two other ends (the B chain and A chain) remain
connected by disulfide bonds.
When insulin was originally purified from bovine or porcine pancreata, all the proinsulin
was not fully removed.[3][4]
When some people used these insulins, the proinsulin may
have caused the body to react with a rash, to resist the insulin, or even to make dents or
lumps in the skin at the place where the insulin was injected. This can be described as
an iatrogenic injury due to slight differences between the proinsulin of different species.
Since the late 1970s, when highly purified porcine insulin was introduced, and the level of
insulin purity reached 99%, this ceased to be a significant clinical issue. The main
challenge for production of insulin using rDNA techniques was getting insulin assembled
into mature form.
Humulin:
Humulin was the first medication produced using modern genetic engineering
techniques in which actual human DNA is inserted into a host cell (E. coli in this case).
Biosynthetic "human" insulin is now manufactured for widespread clinical use using
genetic engineering techniques using recombinant DNA technology, which the
manufacturers claim reduces the presence of many impurities, although there is no
clinical evidence to substantiate this claim. Eli Lilly marketed the first artificial insulin,
Humulin, in 1982.
Humulin production method is as follows:
1. DNA coding for A and B polypeptide chains of insulin are chemically synthesised a
in the lab. Sixty three nucleotides are sequenced to produce A chain of insulin and
ninety nucleotide long DNA designed to produce B chain of insulin, plus terminator
codon is added at the end of each chain sequence. Anti-codon for methionine is
added at the beginning of the sequence to distinguish humulin from the other
bacterial proteins.
2. Chemically synthesized A and B chain DNA sequence are inserted into one of the
marker gene which are present in the plasmid vector. Genes are inserted into the
plasmid with the help of enzymes known as endonuclease and ligase.
3. The vector plasmids with the insulin gene are then introduced into the E. coli
bacterial cell. These cells are then allowed to replicate by mitosis, along with the
bacterial cell recombinant plasmid also gets replicated producing the human
insulin.
4. A and B polypeptide chains of insulin are then extracted and purified from the
fomenters in the lab. High-Performance Liquid Chromatography (HPLC) is used to
get 100% pure humulin from the mixture of proteins.
5. The A and B polypeptide chains of insulin are mixed together and connected with
each other by disulphide bond, forming the Humulin or synthetic human insulin.
Advantages & Disadvantages of Humulin:
Humulin is the one and only human protein produced in the bacteria with identical
chemical structure to that of the natural human insulin. Administration of humulin
reduces the possibility of antibody production and inflammatory response in diabetic
patients. Major difficulty is the
extraction of humulin from a
mixture of host proteins present in
the fermentation broth.
Now days to overcome this
extraction problem synthetic
human insulin are produced in the
yeast cell instead of E. coli using the
same procedure. As yeast is
Eukaryotes they secrete the whole
humulin molecule with perfect three dimensional structures, reducing the need for
complex and time consuming purification methods.
Now most of the diabetic patients are treated with synthetic human insulin. Small group
of patients claim that episodes of hyperglycaemic complications have been increased
after shifting from animal origin insulin to humulin. No study till date shows the
difference between the frequency of hyperglycaemic complications in patient using
humulin (synthetic human insulin) and animal origin insulin.
Gene Therapy
Gene therapy is the therapeutic delivery of nucleic acid polymers into a patient's cells as
a drug to treat disease. Gene therapy is an experimental technique that uses genes to
treat or prevent disease. In the future, this technique may allow doctors to treat a
disorder by inserting a gene into a patient’s cells instead of using drugs or surgery.
Researchers are testing several
approaches to gene therapy,
including:
 Replacing a mutated gene
that causes disease with a
healthy copy of the gene.
 Inactivating, or “knocking
out,” a mutated gene that is
functioning improperly.
 Introducing a new gene into
the body to help fight a
disease.
Although gene therapy is a promising treatment option for a number of diseases
(including inherited disorders, some types of cancer, and certain viral infections), the
technique remains risky and is still under study to make sure that it will be safe and
effective. Gene therapy is currently only being tested for the treatment of diseases that
have no other cures. It should be noted that not all medical procedures that introduce
alterations to a patient's genetic makeup can be considered gene therapy. Bone marrow
transplantation, and organ transplants in general have been found to introduce foreign
DNA into patients. Gene therapy is defined by the precision of the procedure and the
intention of direct therapeutic effects.
Gene therapy was conceptualized in 1972, by authors who urged caution before
commencing human gene therapy studies.
The first attempt, albeit an unsuccessful one, at gene therapy (as well as the first case of
medical transfer of foreign genes into humans not counting organ transplantation) was
performed by Martin Cline on 10 July 1980. Cline claimed that one of the genes in his
patients was active six months later, though he never published this data or had it
verified and even if he is correct, it's unlikely it produced any significant beneficial effects
treating beta-thalassemia.
The first germ line gene therapy consisted of producing a genetically engineered embryo
in October 1996. The baby was born on July 21, 1997 and was produced by taking a
donor's egg with healthy mitochondria, removing its nuclear DNA and filling it with the
nuclear DNA of the biological mother - a procedure known as cytoplasmic transfer.
This procedure was referred to sensationally and somewhat inaccurately in the media as
a "three parent baby", though mtDNA is not the primary human genome and has little
effect on an organism's individual characteristics beyond powering their cells.
Gene therapy is a way to fix a genetic problem at its source. The polymers are
either expressed as proteins, interfere with protein expression, or possibly correct genetic
mutations.
The most common form uses DNA that encodes a functional, therapeutic gene to replace
a mutated gene. The polymer molecule is packaged within a "vector", which carries the
molecule inside cells.
The first commercial gene therapy, Gendicine, was approved in China in 2003 for the
treatment of certain cancers. In 2011 Neovasculgen was registered in Russia as the first-
in-class gene-therapy drug for treatment of peripheral artery disease, including critical
limb ischemia. In 2012 Glybera, a treatment for a rare inherited disorder, became the first
treatment to be approved for clinical use in either Europe or the United States after its
endorsement by the European Commission.
ADA deficiency is one form of SCID (severe combined immunodeficiency), a disorder that
affects the immune system. ADA deficiency is very rare, but very dangerous, because a
malfunctioning immune system
leaves the body open to infection
from bacteria and viruses.
The disease is caused by a mutation
in a gene on chromosome 20. ADA
deficiency is inherited in an
autosomal recessive manner. The
gene codes for the enzyme
adenosine deaminase (ADA).
Without this enzyme, the body is
unable to break down a toxic
substance called deoxyadenosine.
The toxin builds up and destroys
infection-fighting immune cells
called T and B lymphocytes. Because
ADA deficiency affects the immune
system, people who have the
disorder are more susceptible to all kinds of infections, particularly those of the skin,
respiratory system, and gastrointestinal tract. They may also be shorter than normal.
Sadly, most babies who are born with the disorder die within a few months.
Treatments of ADA Deficiency includes:
 bone marrow transplant
 gene therapy
 ADA enzyme in PEG vehicle
On September 14, 1990, the first gene therapy to combat this disease was performed by
Dr. William French Anderson on a four-year-old girl, Ashanti DeSilva, at the National
Institutes of Health, Bethesda, Maryland, U.S.A.
Conclusion
Biotechnology is the new wonder of science. It is truly multidisciplinary in nature and it
encompasses several disciplines of basic sciences and engineering. The Science disciplines
from which biotechnology draws heavily are microbiology, chemistry, biochemistry,
genetics, molecular biology, immunology, cell and tissue culture and physiology. On the
engineering side it leans heavily on process chemical and biochemical engineering since
large scale cultivation of microorganisms and cells, their downstream processing are
based on them. It comes to us as a great blessing...
Biotechnology utilizes the technique called genetic engineering or recombinant DNA
technology where a microorganism is isolated; its genetic material is cut, manipulated,
sealed, again inserted in an organism and allowed to grow in a suitable environment
under controlled conditions to get the desired product. It looks easy but is a very tedious
job and it takes years for a research to achieve its goal.
Like every other thing, biotechnology too has some harmful impacts:
1. Genetic engineering is a very vital part of biotechnology and the cost of
transferring genes from one species to another is very expensive, which requires a
huge amount of capital investment. The cost of producing genetically- modified
plants and animals are sky- rocketing and the duration of return are also not
predictable.
2. Genetic engineering crosses boundaries of reproduction by crossing genes of
species that are completely unrelated; hence giving rise to hazardous results as
well as also increasing the risk of harming multiple species.
3. When genetic material from certain viruses is used in the production of transgenic
crops, there are chances that these virus genes will combine with crop genes to
produce more destructive viruses. The consumption of such crops is hazardous to
human health and can cause several life- threatening ailments. It can also result in
cancer, often malignant as well.
4. Biotechnology also poses a number of environmental threats. Genetically modifies
crops often infect monarch butteries and other insect species.
The applications of biotechnology are so broad, and the advantages so compelling, that
virtually every industry is using this technology. Developments are underway in areas as
diverse as pharmaceuticals, diagnostics, textiles, aquaculture, forestry, chemicals,
household products, environmental cleanup, food processing and forensics to name a
few. Biotechnology is enabling these industries to make new or better products, often
with greater speed, efficiency and flexibility. Biotechnology must continue to be carefully
regulated so that the maximum benefits are received with the least risk.
Bibliography
http://en.wikipedia.org/biotechnology
http://en.wikipedia.org/insulin
http://www.genewatch.org/sub-568238
http://en.wikipedia.org/humulin
http://www.biotecharticles.com/Others-Article/Human-Insulin-and-
Recombinant-DNA-Technology-70.html
https://isaaa.org/resources/publications/pocketk/34/default.asp
http://www.sciencedirect.com/
https://en.wikipedia.org/wiki/Gene_therapy
https://en.wikipedia.org/wiki/Adenosine_deaminase_deficiency
http://www.diabetes.co.uk/insulin/animal-insulin.html
Biology textbook (N.C.E.R.T) Class 12th
Contents
Introduction
History
Biotechnology in Agriculture
 Genetically Modified Crops
 RNA Interference (RNAi)
 Bt toxin
 Bt cotton
Biotechnology in Medicine
 Genetically engineered insulin (Humulin)
 Gene therapy
Conclusion
Bibliography

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294196410-investigatoy-project-on-application-of-biotechnology.docx

  • 1. An English Medium Co. Ed. Senior Secondary School Investigatory Project On S S SU U UB B BM M MI IIT T TT T TE E ED D D B B BY Y Y: : : S S SU U UB B BM M MI IIT T TT T TE E ED D D T T TO O O: : : S S Su u ub b bh h ha a ag g g S S Si iin n ng g gh h h M M Mr r r. .. S S Sa a an n nd d de e ee e ep p p K K Ku u ul lls s sh h he e es s st t th h hr r ra a a X X XI III II S S Sc c ci ii. .. B B B ( ( (H H H. ..O O O. ..D D D B B Bi iio o ol llo o og g gy y y) ) )
  • 2. Aknowledgement I am overwhelmed in all humbleness and gratefulness to acknowledge my depth to all those who have helped me to put these ideas, well above the level of simplicity and into something concrete. I would like to express my special thanks of gratitude to my biology teacher, Mr. Sandeep Kulshesthra as well as our Principal Mrs. Nidhi Bhatia who gave me the golden opportunity to do this wonderful project on the topic “Applications of Biotechnology”, which also helped me in doing a lot of research and I came to know about so many new things. I am really thankful to them. Any attempt at any level can’t be satisfactorily completed without the support and guidance of my Parents and Friends who helped me a lot in gathering different information, collecting data and guiding me from time to time in making this project, despite of their busy schedules, they gave me different ideas in making this project unique. I am thankful to them too. I am making this project not only for marks but to also increase my knowledge... Thanking you Subhag Singh XII Sci. B
  • 3. Certificate This is to certify that SUBHAG SINGH of class XII SCI.B of GYAN DEEP SHIKSHA BHARATI has successfully completed the investigatory project on the topic “APPLICATIONS OF BIOTECHNOLOGY” under the guidance of MR. SANDEEP KULSHESTHRA (H.O.D. Biology) during the session 2015- 16 in the partial fulfilment of Biology Practical Examination conducted by CENTRAL BOARD OF SECONDARY EDUCATION (AISSCE). ___________________ ___________________ Mr. Sandeep Kulshesthra External Examiner (H.O.D Biology) (C.B.S.E)
  • 4. Introduction What is Biotechnology? Biotechnology is the use of living systems and organisms to develop or make products, or "any technological application that uses biological systems, living organisms or derivatives thereof, to make or modify products or processes for specific use. At its simplest, biotechnology is technology based on biology - biotechnology harnesses cellular and bio molecular processes to develop technologies and products that help improve our lives and the health of our planet. We have used the biological processes of microorganisms for more than 6,000 years to make useful food products, such as bread and cheese, and to preserve dairy products. Modern biotechnology provides breakthrough products and technologies to combat debilitating and rare diseases, reduce our environmental footprint, feed the hungry, useless and cleaner energy, and have safer, cleaner and more efficient industrial manufacturing processes. Biotech is helping to heal the world by harnessing nature's own toolbox and using our own genetic makeup to heal and guide lines of research by:  Reducing rates of infectious disease  Saving millions of children's lives  Changing the odds of serious, life-threatening conditions affecting millions around the world  Tailoring treatments to individuals to minimize health risks and side effects  Creating more precise tools for disease detection  Combating serious illnesses and everyday threats confronting the developing world.
  • 5. History 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. 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 mould Penicillium. His work led to the purification of the antibiotic compound formed by the mould 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 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.
  • 6. Biotechnology in Agriculture Genetically Modified Crops Genetically modified crops or “GM crops” or “biotech crops” are plants used in agriculture, the DNA of which has been modified with genetic engineering techniques. In most cases the aim is to introduce a new trait to the plant which does not occur naturally in the species. Examples in food crops include resistance to certain pests, diseases, stressful environmental conditions, resistance to chemical treatments, reduction of spoilage, or improving the nutrient profile of the crop. Examples in non-food crops include production of pharmaceutical agents, bio fuels, and other industrially useful goods,as well as for bioremediation. Plants and crops with GM traits have been tested more than any other crops—with no credible evidence of harm to humans or animals. In fact, seeds with GM traits have been tested more than any other crops in the history of agriculture – with no credible evidence of harm to humans or animals. Governmental regulatory agencies, scientific organizations and leading health associations worldwide agree that food grown from GM crops is safe to eat. The World Health Organization, the American Medical Association, the U.S. National Academy of Sciences, the British Royal Society, among others that have examined the evidence, all come to the same conclusion: consuming foods containing ingredients derived from GM crops is safe to eat and no riskier than consuming the same foods containing ingredients from crop plants modified by conventional plant improvement techniques. Genetic modifications have: 1. Made crops more tolerant to abiotic stresses (cold, drought, salt, heat). 2. Reduced reliance on chemical pesticides (pest resistant crops). 3. Helped to reduce post harvest losses & enhanced the nutritional value of the food.
  • 7. RNA Interference (RNAi) RNA interference (RNAi) is a method of blocking gene function by inserting short sequences of ribonucleic acid (RNA) that match part of the target gene’s sequence, thus no proteins are produced. RNAi has the potential to become a powerful therapeutic approach toward targeted and personalized medicine. RNAi has provided a way to control pests and diseases, introduce novel plant traits and increase crop yield. Using RNAi, scientists have developed novel crops such as nicotine-free tobacco, non-allergenic peanuts, decaffeinated coffee, and nutrient fortified maize among many others. Mechanism of RNA interferences as understood is that it comes into play when a double stranded RNA is introduced either naturally or artificially in a cell. An endo ribonuclease enzyme cleaves the long dsRNA into small pieces of RNA. The small pieces could be mi RNA or si RNA depending upon the origin of long dsRNA i.e. endogenous or exogenous respectively. A double stranded RNA may be generated by either RNA dependent RNA polymerase or bidirectional transcription of transposable elements or physically introduced. There are several opportunities for the applications of RNAi in crop science for its improvement such as stress tolerance and enhanced nutritional level.This knockdown technology may be useful in inducing early flowering, delayed ripening, delayed senescence, breaking dormancy, stress-free plants, overcoming self-sterility, etc. RNA interference (RNAi) has recently been demonstrated in plant parasitic nematodes. It is a potentially powerful investigative tool for the genome-wide identification of gene function that should help improve our understanding of plant parasitic nematodes. RNAi should help identify gene and, hence, protein targets for nematode control strategies. Prospects for novel resistance depend on the plant generating an effective form of double-stranded RNA in the absence of an endogenous target gene without detriment to itself. These RNA molecules must then become available to the nematode and be capable of ingestion via its feeding tube. If these requirements can be met, crop resistance could be achieved by a plant delivering a dsRNA that targets a nematode gene and induces a lethal or highly damaging RNAi effect on the parasite.
  • 8. Bt toxin A protein that is toxic to chewing insects and is produced by the soil bacterium Bacillus thuringiensis and has long been used as a biological pesticide. By means of genetic engineering, the genes for Bt toxin can be isolated from Bacillus thuringiensis and transferred to plants. Bacillus thuringiensis (Bt) is a bacteria that produces proteins which are toxic to insects. But extreme toxicity comes at no surprise. It’s in the same family of bacteria as B. anthracis, which causes anthrax, and B. cereus, which causes food poisoning. The Bt toxin dissolve in the high pH insect gut and become active. The toxins then attack the gut cells of the insect, punching holes in the lining. The Bt spores spills out of the gut and germinate in the insect causing death within a couple days. Even though the toxin does not kill the insect immediately, treated plant parts will not be damaged because the insect stops feeding within hours. Bt spores do not spread to other insects or cause disease outbreaks on their own. 1. Insect eats Bt crystals and spores. 2. The toxin binds to specific receptors in the gut and the insects stops eating. 3. The crystals cause the gut wall to break down, allowing spores and normal gut bacteria to enter the body. 4. The insect dies as spores and gut bacteria proliferate in the body. Bt action is very specific. Different strains of Bt are specific to different receptors in insect gut wall. Bt toxicity depends on recognizing receptors, damage to the gut by the toxin occurs upon binding to a receptor. Each insect species possesses different types of receptors that will match only certain toxin proteins, like a lock to a key. It is because of this that farmers have to be careful to match the target pest species with a particular Bt toxin protein which is specific for that insect. This also helps the benifical insects because they will usually not be harmed by that particular strain of Bt.
  • 9. Bt Cotton Bt cotton is a genetically modified organism (GMO) cotton variety, which produces an insecticide to bollworm. Strains of the bacterium Bacillus thuringiensis produce over 200 different Bt toxins, each harmful to different insects. Most notably, Bt toxins are insecticidal to the larvae of moths and butterflies, beetles, cotton bollworms and ghtu flies but are harmless to other forms of life. The gene coding for Bt toxin has been inserted into cotton as a transgene, causing it to produce this natural insecticide in its tissues. In many regions, the main pests in commercial cotton are lepidopteran larvae, which are killed by the Bt protein in thegenetically modified cotton they eat. This eliminates the need to use large amounts of broad-spectrum insecticides to kill lepidopteran pests. This spares natural insect predators in the farm ecology and further contributes to non insecticide pest management. Bt cotton is ineffective against many cotton pests such as plant bugs, stink bugs, and aphids; depending on circumstances it may be desirable to use insecticides in prevention. A 2006 study done by Cornell researchers, the Center for Chinese Agricultural Policy and the Chinese Academy of Science on Bt cotton farming in China found that after seven years these secondary pests that were normally controlled by pesticide had increased, necessitating the use of pesticides at similar levels to non-Bt cotton and causing less profit for farmers because of the extra expense of GM seeds. Mechanism: Bt cotton was created through the addition of genes encoding toxin crystals in the Cry group of endotoxin. When insects attack and eat the cotton plant the Cry toxins are dissolved due to the high pH level of the insects stomach. The dissolved and activated Cry molecules bond to cadherin-like proteins on cells comprising the brush border molecules. The epithelium of the brush border membranes separates the body cavity from the gut whilst allowing access for nutrients. The Cry toxin molecules attach themselves to specific locations on the cadherin-like proteins present on the epithelial cells of the midge and ion channels are formed which allow the flow of potassium. Regulation of potassium concentration is essential and, if left unchecked, causes death of cells. Due to the formation of Cry ion channels sufficient regulation of potassium ions is lost and results in the death of epithelial cells. The death of such cells creates gaps in the brush border membrane.
  • 10. Advantages: Bt cotton has several advantages over non Bt cotton. The important advantages of Bt cotton are briefly :  Increases yield of cotton due to effective control of three types of bollworms, viz. American, Spotted and Pink bollworms.  Insects belonged to Lepidoptera (Bollworms) are sensitive to crystalline endotoxic protein produced by Bt gene which in turn protects cotton from bollworms.  Reduction in pesticide use in the cultivation of Bt cotton in which bollworms are major pests.  Reduction in the cost of cultivation and lower farming risks.  Reduction in environmental pollution by the use of insecticides rarely.  Bt cotton exhibit genetic resistance or inbuilt resistance which is a permanent type of resistance and not affected by environmental factors. Thus protects crop from bollworms.  Bt cotton is ecofriendly and does not have adverse effect on parasites, predators, beneficial insecticides and organisms present in soil.  It promotes multiplication of parasites and predators which help in controlling the bollworms by feeding on larvae and eggs of bollworm.  No health hazards due to rare use of insecticides.  Bt cotton are early in maturing as compared to non Bt cotton. Disadvantages: Bt cotton has some limitations  High cost of Bt cotton seeds as compared to non Bt cotton seeds.  Effectiveness up to 120 days, after that the toxin producing efficiency of the Bt gene drastically reduces.  Ineffective against sucking pests like jassids, aphids, whitefly etc.
  • 11. Bt cotton in India: Bt cotton is supplied in India's Maharashtra state by the agri-biotechnology company, Mahyco, as the distributor. The use of Bt cotton in India has grown exponentially since its introduction. Recently India has become the number one global exporter of cotton and the second largest cotton producer in the world. India has bred Bt-cotton varieties such as Bikaneri Nerma and hybrids such as NHH-44, setting up India to benefit now and well into the future. India’s success has been subject to scrutiny. Monsanto's seeds are expensive and lose vigour after one generation, prompting the Indian Council of Agricultural Research to develop a cheaper Bt cotton variety with seeds that could be reused. The cotton incorporated the cry1Ac gene from the soil bacterium Bacillus thuringiensis (Bt), making the cotton toxic to bollworms. In parts of India cases of acquired resistance against Bt cotton have occurred. The state of Maharashtra banned the sale and distribution of Bt cotton in 2012, to promote local Indian seeds, which demand less water, fertilizers and pesticide input, but lifted the ban in 2013. India approved Bt cotton in 2002; now it accounts for 92% of all Indian cotton. Average nationwide cotton yields went from 302 kg/ha in the 2002/3 season to a projected 481 kg/ha in 2011/12 — up 59.3% overall. This chart shows the trends in yields, which took off after Bt was introduced in 2002. The graphs also show that — and here comes ugly fact— in the last 4 years, as Bt has risen from 67% to 92% of India’s cotton, yields have dropped steadily.
  • 12. Biotechnology in Medicine Genetically Engineered Insulin (Humulin) Insulin is a peptide hormone produced by beta cells in the pancreas of various organisms including human beings. It regulates the metabolism of carbohydrates and fats by promoting the absorption of glucose from the blood to skeletal muscles and fat tissue and by causing fat to be stored rather than used for energy. Insulin also inhibits the production of glucose by the liver. Except in the presence of the metabolic disorder diabetes mellitus and metabolic syndrome, insulin is provided within the body in a constant proportion to remove excess glucose from the blood, which otherwise would be toxic. When blood glucose levels fall below a certain level, the body begins to use stored glucose as an energy source through glycogenolysis, which breaks down the glycogen stored in the liver and muscles into glucose, which can then be utilized as an energy source. As a central metabolic control mechanism, its status is also used as a control signal to other body systems (such as amino acid uptake by body cells). In addition, it has several other anabolic effects throughout the body. When control of insulin levels fails, diabetes mellitus can result. Structure: Insulin is composed of two different types of peptide chains. Chain A has 21 amino acids and Chain B has 30 amino acids. Both chains contain alpha helices but no beta strands. There are 3 conserved disulfide bridges which help keep the two chains together. Insulin can also form dimers in solution due to the hydrogen bonding between the B chains. The dimers can further interact
  • 13. to form hexamers due to interaction between hydrophobic surfaces. This scene highlights the hydrophobic and polar parts of an insulin monomer at a pH of 7. A number of insulin variants have been made to favor either the monomeric or hexameric form. Deletion of the five C terminal residues of the B chain creates a monomer only form. This portion of the B chain is involved in hydrogen bonds between the B chain of one monomer and the A and B chain of another monomer. Need of Genetically Engineered Insulin: The original form of the wonder cure for diabetes, these were once the only type of insulin available, but are now rarely used. Animal insulin was originally made from ground-up animal pancreas tissue, and then later was extracted from healthy animals (slaughtered pigs & cows). The metabolism of cows and pigs was close enough to human metabolism that their animal insulin also worked well in human bodies. Beef insulin has 3 differences from human; pork insulin has 1 difference from human. The use of a mixture of beef and pork insulin was also possible. It has been shown that human insulin is less immunogenic than animal insulin. Porcine insulin is most similar to human insulin. The primary amino acid sequences of bovine and porcine insulin differ from that of human insulin by three and one amino acid, respectively. This greater dissimilarity between human and bovine insulin has been postulated to be the explanation for the greater antigenicity of bovine insulin as compared with porcine insulin One of the problems with animal insulin was antibody issues. The body identifies them and tries to reject them. Pork insulin differs by 1 amino acid and beef insulin by 3 amino acids, so the body's immune system can sometimes recognize them as foreign. Immunological complications of insulin therapy have been evident since animal insulin became available for the treatment of diabetes mellitus in 1922. In insulin-allergic patients treated with conventional insulin preparations, the insulin-specific IgE values are often 10- to 20-fold higher than in patients without allergy. It has been shown that human insulin is less immunogenic than animal insulin. Porcine insulin is most similar to human insulin. Cross-reactivity between human insulin and insulin of animal origin has been reported. A major problem is the cross-reactivity that occurs between anti-insulin antibodies and the various animal and human insulin preparations in patients presenting with allergy to animal insulin.
  • 14. The usage of animal insulin has so greatly declined in modern times that they have largely been withdrawn from the market. Newly diagnosed diabetics are typically given synthesized or Genetically Engineered human insulin. What is “Proinsulin”? Proinsulin is the prohormone precursor to insulin made in the beta cells of the islets of Langerhans, specialized regions of the pancreas. Proinsulin is synthesized on membrane associated ribosomes found on the rough endoplasmic reticulum, where it is folded and its disulfide bonds are oxidized. It is then transported to the Golgi apparatus where it is packaged into secretory vesicles, and where it is processed by a series of proteases to form mature insulin. Mature insulin has 35 fewer amino acids; 4 are removed altogether, and the remaining 31 form the C-peptide. The C-peptide is abstracted from the center of the proinsulin sequence; the two other ends (the B chain and A chain) remain connected by disulfide bonds. When insulin was originally purified from bovine or porcine pancreata, all the proinsulin was not fully removed.[3][4] When some people used these insulins, the proinsulin may have caused the body to react with a rash, to resist the insulin, or even to make dents or lumps in the skin at the place where the insulin was injected. This can be described as an iatrogenic injury due to slight differences between the proinsulin of different species. Since the late 1970s, when highly purified porcine insulin was introduced, and the level of insulin purity reached 99%, this ceased to be a significant clinical issue. The main challenge for production of insulin using rDNA techniques was getting insulin assembled into mature form. Humulin: Humulin was the first medication produced using modern genetic engineering techniques in which actual human DNA is inserted into a host cell (E. coli in this case). Biosynthetic "human" insulin is now manufactured for widespread clinical use using genetic engineering techniques using recombinant DNA technology, which the manufacturers claim reduces the presence of many impurities, although there is no clinical evidence to substantiate this claim. Eli Lilly marketed the first artificial insulin, Humulin, in 1982. Humulin production method is as follows: 1. DNA coding for A and B polypeptide chains of insulin are chemically synthesised a in the lab. Sixty three nucleotides are sequenced to produce A chain of insulin and
  • 15. ninety nucleotide long DNA designed to produce B chain of insulin, plus terminator codon is added at the end of each chain sequence. Anti-codon for methionine is added at the beginning of the sequence to distinguish humulin from the other bacterial proteins. 2. Chemically synthesized A and B chain DNA sequence are inserted into one of the marker gene which are present in the plasmid vector. Genes are inserted into the plasmid with the help of enzymes known as endonuclease and ligase. 3. The vector plasmids with the insulin gene are then introduced into the E. coli bacterial cell. These cells are then allowed to replicate by mitosis, along with the bacterial cell recombinant plasmid also gets replicated producing the human insulin. 4. A and B polypeptide chains of insulin are then extracted and purified from the fomenters in the lab. High-Performance Liquid Chromatography (HPLC) is used to get 100% pure humulin from the mixture of proteins. 5. The A and B polypeptide chains of insulin are mixed together and connected with each other by disulphide bond, forming the Humulin or synthetic human insulin. Advantages & Disadvantages of Humulin: Humulin is the one and only human protein produced in the bacteria with identical chemical structure to that of the natural human insulin. Administration of humulin reduces the possibility of antibody production and inflammatory response in diabetic patients. Major difficulty is the extraction of humulin from a mixture of host proteins present in the fermentation broth. Now days to overcome this extraction problem synthetic human insulin are produced in the yeast cell instead of E. coli using the same procedure. As yeast is Eukaryotes they secrete the whole humulin molecule with perfect three dimensional structures, reducing the need for complex and time consuming purification methods. Now most of the diabetic patients are treated with synthetic human insulin. Small group of patients claim that episodes of hyperglycaemic complications have been increased after shifting from animal origin insulin to humulin. No study till date shows the difference between the frequency of hyperglycaemic complications in patient using humulin (synthetic human insulin) and animal origin insulin.
  • 16. Gene Therapy Gene therapy is the therapeutic delivery of nucleic acid polymers into a patient's cells as a drug to treat disease. Gene therapy is an experimental technique that uses genes to treat or prevent disease. In the future, this technique may allow doctors to treat a disorder by inserting a gene into a patient’s cells instead of using drugs or surgery. Researchers are testing several approaches to gene therapy, including:  Replacing a mutated gene that causes disease with a healthy copy of the gene.  Inactivating, or “knocking out,” a mutated gene that is functioning improperly.  Introducing a new gene into the body to help fight a disease. Although gene therapy is a promising treatment option for a number of diseases (including inherited disorders, some types of cancer, and certain viral infections), the technique remains risky and is still under study to make sure that it will be safe and effective. Gene therapy is currently only being tested for the treatment of diseases that have no other cures. It should be noted that not all medical procedures that introduce alterations to a patient's genetic makeup can be considered gene therapy. Bone marrow transplantation, and organ transplants in general have been found to introduce foreign DNA into patients. Gene therapy is defined by the precision of the procedure and the intention of direct therapeutic effects. Gene therapy was conceptualized in 1972, by authors who urged caution before commencing human gene therapy studies. The first attempt, albeit an unsuccessful one, at gene therapy (as well as the first case of medical transfer of foreign genes into humans not counting organ transplantation) was performed by Martin Cline on 10 July 1980. Cline claimed that one of the genes in his patients was active six months later, though he never published this data or had it verified and even if he is correct, it's unlikely it produced any significant beneficial effects treating beta-thalassemia. The first germ line gene therapy consisted of producing a genetically engineered embryo in October 1996. The baby was born on July 21, 1997 and was produced by taking a donor's egg with healthy mitochondria, removing its nuclear DNA and filling it with the nuclear DNA of the biological mother - a procedure known as cytoplasmic transfer.
  • 17. This procedure was referred to sensationally and somewhat inaccurately in the media as a "three parent baby", though mtDNA is not the primary human genome and has little effect on an organism's individual characteristics beyond powering their cells. Gene therapy is a way to fix a genetic problem at its source. The polymers are either expressed as proteins, interfere with protein expression, or possibly correct genetic mutations. The most common form uses DNA that encodes a functional, therapeutic gene to replace a mutated gene. The polymer molecule is packaged within a "vector", which carries the molecule inside cells. The first commercial gene therapy, Gendicine, was approved in China in 2003 for the treatment of certain cancers. In 2011 Neovasculgen was registered in Russia as the first- in-class gene-therapy drug for treatment of peripheral artery disease, including critical limb ischemia. In 2012 Glybera, a treatment for a rare inherited disorder, became the first treatment to be approved for clinical use in either Europe or the United States after its endorsement by the European Commission. ADA deficiency is one form of SCID (severe combined immunodeficiency), a disorder that affects the immune system. ADA deficiency is very rare, but very dangerous, because a malfunctioning immune system leaves the body open to infection from bacteria and viruses. The disease is caused by a mutation in a gene on chromosome 20. ADA deficiency is inherited in an autosomal recessive manner. The gene codes for the enzyme adenosine deaminase (ADA). Without this enzyme, the body is unable to break down a toxic substance called deoxyadenosine. The toxin builds up and destroys infection-fighting immune cells called T and B lymphocytes. Because ADA deficiency affects the immune system, people who have the disorder are more susceptible to all kinds of infections, particularly those of the skin, respiratory system, and gastrointestinal tract. They may also be shorter than normal. Sadly, most babies who are born with the disorder die within a few months. Treatments of ADA Deficiency includes:  bone marrow transplant  gene therapy  ADA enzyme in PEG vehicle
  • 18. On September 14, 1990, the first gene therapy to combat this disease was performed by Dr. William French Anderson on a four-year-old girl, Ashanti DeSilva, at the National Institutes of Health, Bethesda, Maryland, U.S.A. Conclusion Biotechnology is the new wonder of science. It is truly multidisciplinary in nature and it encompasses several disciplines of basic sciences and engineering. The Science disciplines from which biotechnology draws heavily are microbiology, chemistry, biochemistry, genetics, molecular biology, immunology, cell and tissue culture and physiology. On the engineering side it leans heavily on process chemical and biochemical engineering since large scale cultivation of microorganisms and cells, their downstream processing are based on them. It comes to us as a great blessing... Biotechnology utilizes the technique called genetic engineering or recombinant DNA technology where a microorganism is isolated; its genetic material is cut, manipulated, sealed, again inserted in an organism and allowed to grow in a suitable environment under controlled conditions to get the desired product. It looks easy but is a very tedious job and it takes years for a research to achieve its goal. Like every other thing, biotechnology too has some harmful impacts: 1. Genetic engineering is a very vital part of biotechnology and the cost of transferring genes from one species to another is very expensive, which requires a huge amount of capital investment. The cost of producing genetically- modified plants and animals are sky- rocketing and the duration of return are also not predictable. 2. Genetic engineering crosses boundaries of reproduction by crossing genes of species that are completely unrelated; hence giving rise to hazardous results as well as also increasing the risk of harming multiple species. 3. When genetic material from certain viruses is used in the production of transgenic crops, there are chances that these virus genes will combine with crop genes to produce more destructive viruses. The consumption of such crops is hazardous to human health and can cause several life- threatening ailments. It can also result in cancer, often malignant as well. 4. Biotechnology also poses a number of environmental threats. Genetically modifies crops often infect monarch butteries and other insect species. The applications of biotechnology are so broad, and the advantages so compelling, that virtually every industry is using this technology. Developments are underway in areas as diverse as pharmaceuticals, diagnostics, textiles, aquaculture, forestry, chemicals, household products, environmental cleanup, food processing and forensics to name a few. Biotechnology is enabling these industries to make new or better products, often with greater speed, efficiency and flexibility. Biotechnology must continue to be carefully
  • 19. regulated so that the maximum benefits are received with the least risk. Bibliography http://en.wikipedia.org/biotechnology http://en.wikipedia.org/insulin http://www.genewatch.org/sub-568238 http://en.wikipedia.org/humulin http://www.biotecharticles.com/Others-Article/Human-Insulin-and- Recombinant-DNA-Technology-70.html https://isaaa.org/resources/publications/pocketk/34/default.asp http://www.sciencedirect.com/ https://en.wikipedia.org/wiki/Gene_therapy https://en.wikipedia.org/wiki/Adenosine_deaminase_deficiency http://www.diabetes.co.uk/insulin/animal-insulin.html Biology textbook (N.C.E.R.T) Class 12th
  • 20. Contents Introduction History Biotechnology in Agriculture  Genetically Modified Crops  RNA Interference (RNAi)  Bt toxin  Bt cotton Biotechnology in Medicine  Genetically engineered insulin (Humulin)  Gene therapy Conclusion Bibliography