Professional Documents
Culture Documents
On
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. Y.K.
Saxena 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
Amitesh Singh
XII Science
Certificate
This is to certify that AMITESH SINGH of class XII SCIENCE
of DAV SR. SEC. PUBLIC SCHOOL,ANPARA has successfully
completed the investigatory project on the topic
“APPLICATIONS OF BIOTECHNOLOGY” under the guidance
of MR. Y.K. SAXENA ( Biology Teacher) during the session
2022-23 in the partial fulfilment of Biology Practical
Examination conducted by CENTRAL BOARD OF
SECONDARY EDUCATION (AISSCE).
___________________ ___________________
Mr. Y.K. Saxena External Examiner
( Biology teacher) (C.B.S.E)
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
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
1. Made crops more tolerant to abiotic stresses (cold, drought, salt, heat).
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.
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.
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
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.
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.
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:
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