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APPLICATIONS OF BIOTECHNOLOGY
Applications of Biotechnology
Applications in Agriculture
Genetically Modified Organisms (GMOs): organisms which are genetically engineered for
human benefit.
Genetic modifications of plants have following advantages:
1. Abiotic stress tolerant crops
a. GM plants can tolerate extreme cold, draught, heat, high salt conc. in soil, etc.
2. Reduced reliance on chemical pesticides
a. Reduces the side effects of pollution caused by chemical pesticides
b. Insect resistant and pest resistant plants were developed
3. Reduced post harvest losses
a. GM plants have higher shelf life
b. They are resistant to microbial infections
4. Increased efficiency of mineral usage by plants
a. So that the soil fertility is not exhausted early
5. Enhanced nutritional value
a. So that food security can be ensured
b. Plants rich in vitamins, calcium, etc are created
Bacteria Bacillus thuringiensis contain cry genes – cryIAb, cryIAc and cryIIAb
Cry genes produce insecticidal proteins effective against the cotton infecting insects.
However, they are produced in inactive form inside bacteria and hence do not effect them.
Genetic engineering of cotton plants: the process is as follows
They reduce the plants roots’ absorbing capacity and thus reduce the yield.
A novel strategy known as RNA interference or RNA Silencing is employed to make pest
resistant plants.
30 therapeutic recombinant proteins are approved for human use, 12 are marketed in India.
One of the most common diseases among all human is Diabetes.
Since the time it was discovered that deficiency of hormone Insulin is one important reason
for its onset, different insulin molecules from various sources such as cow and pig have been
tried.
Although, they were functional to some extent and induces immunogenic response in the
body.
After the birth of genetic engineering, recombinant human insulin is being made in a
bacterial host.
The first such recombinant human insulin is branded as Humulin
It was manufactured by Eli Lilly and Co., and American company in 1983.
Structure of normal in vivo Human Insulin
Gene Therapy
Gene therapy literally means replacing the defective gene by a functional copy of the same.
First gene therapy was successfully carried out for ADA-SCID in a 4 year old Sri Lankan girl
Ashanti de-Silva by Dr. William French Anderson at National Institute of Health, USA in
1990
ADA-SCID: Adenosine Deaminase Severe Combined Immunodeficiency
o Adenosine is an enzyme coded by ada gene in chromosome 20
o It is expressed in the lymphocytes necessary for proper immune system development
o Its deficiency causes severyly compromised immune system
o Conventional treatment includes –
Enzyme Replacement Therapy – not a permanent treatment procedure as
frequent injections of enzyme is necessary
Bone Marrow Transplantation – not feasible always due to non availability of
matching donor
Gene Therapy for ADA SCID
o Since lymphocytes have a fixed life span, GM lymphocytes need to regularly injected
into patient’s blood.
o This limitation may be removed by carrying out gene therapy at an early embryonic
stage so that all the cells of body receive the normal copy of the gene.
Molecular Diagnosis
Many acute diseases such as AIDS and Cancer require early detection to be treated
efficiently.
However, their diagnosis depends upon the quantity of infectious agents or amount of
mutations present in the body.
Thus PCR is employed to detect the presence of bacteria or virus or mutations in DNA
Process of detection
o Primers are used to amplify the pathogen DNA or
the mutated DNA
o If the pathogen is present even in very minute
quantity, its DNA is amplified
o During the electrophoresis, a band for pathogen
DNA is observed along with the host DNA.
Types of PCR for diagnosis –
o Qualitative PCR – only presence or absence of a
pathogen is detected
o Quantitative PCR – quantity of pathogen or
mutation is detected.
↓
A substrate is added for the enzyme which changes colour
when acted upon by the enzyme thus showing the presence of
antigen in the serum
↓
Qualitative Quantitative
estimate estimate
Probe: an artificially synthesized single stranded DNA or RNA sequence of variable length
(usually 100-1000 bases long).
A radiolabelled probe has a radioactive substance coupled to it.
The sequence of the probe is complementary to the target DNA sequence to be identified.
This technique can be used for identifying the presence of mutations in a sample of tissues.
A brief overview of the principle:
A radiolabelled probe is synthesized having sequence complementary to normal gene
sequence
↓
The DNA from tissue sample is exposed to Radiolabelled probe for hybridisation
↓
Hybridised sample is exposed to X ray film for detecting radioactive signals
↓
Interpretation of observations:
If the gene is not mutated, probe binds to it and gives a positive signal for radioactivity
If the gene is mutated, its sequence changes, as a result, probe does not bind to it, and gives a
negative result
TRANSGENIC ANIMALS
to study normal
physiology and
development
to study to study a
chemical human
toxicity disease and
new drug
TRANSGENIC trial
ORGANISMS
to produce
To study the useful
vaccine safety biological
products
Model organism: an organism that has a physiology similar to human beings, which is used
to study human physiology or pathophysiology.
o Transgenic Model Organisms: they are produced by introducing a foreign gene
into a model organism which can produce a typical phenotype similar to human.
o Knock out model organism: a transgenic animal in which a specific gene is
inactivated by introducing another similar gene (having enough changes in sequences
to produce an abnormal copy of the gene product)
The introduced gene due to its homology with the host gene gets incorporated in the
place of host gene by replacing the same.
o E.g. - model organisms are available for cancer, cystic fibrosis, rheumatoid arthritis,
Alzheimer’s disease, etc.
Production of Biological Products:
o Various biological products have uses in medicine.
o But they are either difficult to extract from animal tissues or difficult or too expensive
to synthesize.
o Such products may be produced by genetic engineering as follows:
o E.g. –
1. 1997, 1st transgenic cow, Rosie was born, produced 2.4 gm/ltr of α-lactalbumin
2. Human protein α-1-antitrypsin for treatment of emphysema
3. Tissue Plasminogen Activator from transgenic goat
4. Blood Clotting Factor VIII and IX from transgenic sheep
ETHICAL ISSUES
Ethics: ethics is a set of moral principles and standards by which a community regulates its
behaviour and decides as to the legitimacy and illegitimacy of an activity.
Bioethics: a set of moral principles and standards which regulate our perceptions as to
exploitation of biological world
Biopatent
Patent: an official right from a government granting an inventor or establishment the sole
monopoly to make and sell a particular article for a certain period.
Biopatent: an official right from a government granting an inventor or establishment the sole
monopoly to use a particular biological material for commercial exploitation of
i. Strains of microorganisms
ii. Cell lines
iii. Genetically modified plants and animals
iv. DNA sequences
v. Protein encoded by DNA sequences
vi. A particular biotechnological procedure
vii. A production process
viii. Product
ix. Product application
Ethical issues related to biopatenting: Cases study
o Certain developed countries are issuing broad patents for whole group of organisms
which prevent others from using any related organism or for carrying out any
research.
o Case of Basmati rice:
27 documented varieties of Basmati rice are grown in India.
An American company developed a new variety of basmati by crossing an Indian
farmer’s variety and semi dwarf variety.
The company got patent for the new variety from US patent and Trademark
office, which allowed the company to sell the new variety in the US and abroad.
The US patent extends to functional equivalents which mean it prevents anyone
else from selling Basmati rice.
Biopiracy
Action taken
Indian Parliament has amended the Indian Patent Bill.
It now considers emergency granting of patent for research and development initiative.