Professional Documents
Culture Documents
Investigatory Project On: An English Medium Co. Ed. Senior
Investigatory Project On: An English Medium Co. Ed. Senior
Senior
Secondary School
Investigatory
Project On
SUBMITTED BY:
SUBMITTED TO:
Subhag Singh Mr.
Sandeep Kulshesthra
XII Sci. B
(H.O.D Biology)
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
(C.B.S.E)
(H.O.D Biology)
Introduction
What is Biotechnology?
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.
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.
1. Made crops more tolerant to abiotic stresses (cold, drought, salt, heat).
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.
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.
Disadvantages:
Bt cotton has some limitations
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 an d 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.
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.
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.
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
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
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.
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.
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:
gene therapy
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:
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_deficie
ncy
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