12

BIOTECHNOLOGY
AND
IT APPLICATION
KEYNOTES 2.0
By: Dr. Anand Mani

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Biotechnology has a wide range application such as biopharmaceuticals,therapeutics, diagnostics, genetically
modified crops for agriculture, processed food, bioremediation, waste treatment and energy production.

BIOTECHNOLOGY HAS 3 CRITICAL RESEARCH AREAS:

1 2
Providing the Downstream

3
best catalyst in Creating optimal processing
the form of conditions through technologies to
improved engineering purify
organism usually for a catalyst to the
a microbe or act. protein/organic
pure enzyme. compound.

APPLICATIONS IN AGRICULTURE
3 OPTIONS FOR INCREASING FOOD PRODUCTION:

• Agro-chemical based • Organic agriculture. • Genetically

agriculture. engineered crop-based
agriculture.

Genetically Modified Organisms (GMO) or transgenic organisms are the plants, bacteria, fungi & animals
whose genes are altered bymanipulation

Advantages of genetic modification in plants:

It makes crops more tolerant to abiotic stresses (cold, drought, salt, heat etc).
Pest-resistant crops reduce the use of chemical pesticides. It helps to reduce post-harvest
losses.

It increases efficiency of mineral usage by plants (it prevents early exhaustion of fertility
of soil). It enhances nutritional value of food. E.g. Vitamin enriched rice. GOLDENRICE
To create tailor-made plants to supply alternativeresources (starches, fuels,
pharmaceuticals etc.) to industries

MEDICINE HIRUDIN
PEST RESISTANTPLANTS BrassicaNapus
• They act as • It reduces the • E.g. Bt cotton, Bt corn,
bio-pesticide. need for insecticides. rice, tomato, potato,
soyabean etc.

Flaur SAUR Polygalacturinase

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BT COTTON:

Some strains of Bacillus thuringiensis have proteins that kill insects like coleopterans
lepidopterans (tobaccobudworm, armyworm) & dipterans (flies, mosquitoes).

B. thuringiensis forms a toxic insecticidal protein (Bt toxin) crystal during a particular
phase of their growth. It does not kill the Bacillus as it exists as inactive protoxins.

When an insect ingests the toxin, it becomes active due to the alkaline pH of the gut
which solubilise the crystals. The toxin binds to the surface of mid-gut epithelial cells and
creates pores. It causes cell swelling and lysis and death of the insect.

Bt toxin genes were isolated from B. thuringiensis and incorporated into crop plants
such as cotton.

Most Bt toxins are insect-group specific. The toxin is coded by a gene named cry.
E.g. proteins encoded by the genes cryIAc & cryIIAb control the cotton bollworms that
of cryIAb controls corn borer.

Nematode resistance in tobacco plants:

A nematode Meloidogyne incognitia infects the roots of tobacco plants causing a

reduction in yield.

It can be prevented by RNA interference (RNAi) strategy.

RNAi is a method of cellular defense in all eukaryotic organisms. It prevents translation

of a specific mRNA (silencing) due to a complementary dsRNA molecule.
C

The source of this complementary RNA is from an infection by RNA viruses or mobile
genetic elements (transposons) that replicate via an RNA intermediate.

Using Agro bacterium vectors, nematode-specific genes (DNA) are introduced into host
plant. It produces both sense & anti-sense RNA in host cells. These RNAs are
complementary. So they form double stranded (ds) RNA. It initiates RNAI and silences the
specific mRNA of nematode. Thus the parasite cannot survive in a transgenic host
expressing specific interfering RNA.

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 APPLICATIONS INMEDICINE
The products from non-human sources
induce unwanted immunological responses.
But recombinant therapeutics does not
have such problems.
The recombinant DNA technology
helps for the mass production of safe
and more effective therapeutic drugs. At present, about 30 recombinant
therapeutics have been approved. Of
these, 12 are being marketed in India.

1. Genetically Engineered Insulin

Insulin is used to manage adult-onset diabetes.

Insulin from the pancreas of animals (cattle and pigs) causes allergy or
other types of reactions to the foreign protein.

Now, it is possible to produce human insulin using bacteria.

Insulin consists of two short polypeptide chains (chain A & chain B)

that are linked by disulphide bridges.

In mammals, insulin is synthesized as a pro hormone. It needs become

(pro- insulin). Processing to mature and functional hormone.

The pro- hormone contains an extra stretch called C peptide. This is

removed during maturation into insulin.

In 1983, Eli Lilly (an American company) prepared two DNA sequences
corresponding to A & B chains of human insulin and introduced them in
plasmids of E. coli to produce insulin chains. Chains A & B were
combined by creating disulfide bonds to form human insulin.

2. Gene Therapy SD
It is a method to correct a gene defect in a Here, genes are inserted into a person’s cells
child/embryo. and tissues to treat a hereditary disease. It
compensates for the non functional gene.

First clinical gene therapy (1990) was given to

a 4-year Old girl with adenosine deaminase This can be cured by bone marrow
(ADA) deficiency. The disorder is caused due to transplantation or by enzyme replacement
the deletion of the gene for adenosine deam therapy (injection of functional ADA). But
inase (an enzyme crucial for the immune these approaches are not completely
system to function). curative.

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In gene therapy, lymphocytes from the patient’s blood are grown in a culture. Then, a functional
ADA cDNA (using a retroviral vector) is introduced into these lymphocytes. Then, they are
returned to the patient. This should be periodically repeated as these cells are not immortal. If
the ADA gene (from marrow cells) is introduced into cells at early embryonic
stages, it could be a permanent cure.

3. MOLECULAR DIAGNOSIS
Early diagnosis of diseases using conventional methods (serum and urine analysis)
are not possible.

Recombinant DNA technology, PCR & ELISA are some techniques for early diagnosis.

PCR (POLYMERASE CHAIN REACTION):

Presence of a pathogen is normally However, very low concentration

suspected only based on symptoms. of a bacteria or virus can be
By this time, the concentration of detected by amplification of their
pathogen is already very high in the nucleic acid by PCR.
body.
Probe
USES OF PCR:
- To detect HIV in suspected AIDS patients.
- To detect gene mutations in suspected cancer patients.
- To identify many other genetic disorders.
- A single stranded DNA or RNA, tagged with a radioactive molecule (probe) is hybridized to its
complementary DNA in a clone of cells followed by detection using autoradiography. The clone
having mutated gene will not appear on the photographic film, because the probe will not
have complementarity with the mutated gene.

ELISA (ENZYME LINKED IMMUNO-SORBENT ASSAY):

- It is based on the principle of antigen-antibody interaction.
- Infection by pathogen can be detected by the presence of antigens (proteins, glycoproteins,
etc.) or by detecting the antibodies synthesized against the pathogen.

western blottingtechnique
TRANSGENIC ANIMALS
- These are the animals whose genome has been altered by introduction of an extra (foreign)
gene by manipulation.
- E.g. Transgenic rats, rabbits, pigs, sheep, cows and fish.
- Over 95% of all existing transgenic animals are mice.

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 BENEFITS OF TRANSGENIC ANIMALS

To study the regulation of genes and

To Study the contribution of genes in the
their action on normal physiology &
development of a disease and thereby new
development:
treatments: E.g. transgenic models for many
E.g. study of complex factors such as
human diseases such as cancer, cystic fibrosis,
insulin-like growth factor. Genes (from
rheumatoid arthritis & Alzheimer’s. Biological
other species) that alter the formation
products: Some medicines contain expensive
of this factor are introduced and the
biological products. Transgenic animals are
biological effects are studied. This gives
used to produce useful biological products by
information about the biological role of
introducing genes which codes for a particular
the factor in the body.
product.

E.g. human protein (a-1-antitrypsin) used to treat emphysema, products for treatment of
phenylketonuria (PKU) and cystic fibrosis etc. In 1997, Rosie (first transgenic cow) produced
human protein-enriched milk (2.4 gm per litre). It contains the human
a -lactalbumin and is nutritionally more balanced product for human babies than natural cowmilk.
Vaccine safety testing: Transgenic mice are used to test the safety of the polio vaccine. If it is
reliable, they can replace the use of monkeys to test the safety of
vaccines. Chemical safety testing (toxicity testing): Transgenic animals are made that carry
genes which make them more sensitive to toxic substances than non-transgenic animals. They
are exposed to the toxic substances and the effects studied. It gives immediate results.

ETHICAL ISSUES
E.g. Basmati rice, herbal medicines
(turmeric, neem etc). Basmati rice
Problem of unpredictable results: has unique aroma & flavour. India
Genetic modification may cause has 27 varieties of Basmati. In 1997,
unpredictable results. an American company got
Indian Government has set up patent rights on Basmati rice
organizations like GEAC (Genetic through the US Patent and
Engineering Approval Committee) to Trademark Office. This
make decisions about the validity of GM allowed the company to sell a
research and the safety of GM- ‘new’ variety of Basmati.
organisms for public services. Problems
of patent: Certain companies have got
patents for products and technologies This was actually derived from
that make use of the genetic materials, Indian farmer’s varieties. Indian
plants etc. that have been identified, Basmati was crossed with semi-
developed and used by farmers and dwarf varieties and claimed as a
indigenous people of a country. novelty. Other people selling
Basmati rice could be restricted
by patent.

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Biopiracy:

It is the use of bio-resources by multinational companies and other

organizations without proper authorization from the countries and people
concerned. Most of the industrialized nations are poor in biodiversity and
traditional knowledge. The developing and the underdeveloped world have
rich biodiversity and traditional knowledge related to bio-resources. It has to
develop laws to prevent unauthorized exploitation of bio-resources and
traditional knowledge. Indian Parliament has cleared the second amendment
of the Indian Patents Bill that has considered patent terms emergency
provisions and research and development initiative.

additional points

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