BIOLOGY INVESTIGATORY PROJECT

ON

BIOTECHNOLOGY AND ITS

APPLICATIONS

SUBMITTED BY: Abhishek bhati

12th A

SUBMITTED TO: Daya chaturvedi

(H.O.D BIOLOGY)
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 (RNA i)

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 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
imunominogenic 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.

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 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 firstin-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
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-
andRecombinant-DNA-Technology-70.html

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

Biologytextbook (N.C.E.R.T) Class 12th