Chapter Outline.

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BIOTECHNOLOGY AND ITS APPLICATIONS

➢ Biotechnology applications-
- Industrial scale production of biopharmaceuticals
- genetically modified crops for agriculture
- therapeutics
- diagnostics
- processed food
- bioremediation
- waste treatment
- energy production
- GM animals, microbes, fungi, plants

➢ Critical research areas of Biotech-

- Providing best catalyst in form of microbe or pure enzyme
- Creating optimal conditions – catalyst to act
- Downstream processing technologies

➢ Application in agriculture:
- Green Revolution- tripled the food supply. Not enough though.

- Genetically modified plants has:

• made crops more tolerant – abiotic stresses (cold, drought, salt,
heat)
• reduced reliance on chemical pesticides (pest- resistant crops)
• reduced post-harvest loss
• increased efficiency of mineral usage by plants (prevents early
exhaustion of fertility of soil)
• enhanced nutritional value of food (golden rice – vitamin A
enriched rice)
 - GM has been used to made tailor made plants- resources to industries
(starches, fuels and pharmaceuticals)
➢ Bt cotton:
produced by a bacterium called Bacillus thuringiensis
cloned from the bacteria -> expressed in plants -> resistance to insects
(without insecticides); in effect created bio-pesticide.
Examples: Bt cotton, Bt corn, rice, tomato, potato and soyabean
During growth phase- Bt forms toxic insecticidal protein
Toxin exists in inactive form- ingested my insect- alkaline pH of gut- active
toxin- death of insect.
Choice of Bt toxin genes is isolated depends on- crop and targeted pest
Toxin coded by – cry gene

➢ Pest- resistant plant-

Nematode Meloidegyne incognitia affect roots of tobacco
Prevent infestation -RNA interference strategy
RNAi- method of self defence (eukaryotic organisms)
Specific gene (DNA) that produce both sense and anti-sense RNA-
introduced in host plant
Complementary RNA form dsRNA that initiates RNAi and allowed
silencing for specific m-RNA of nematodes.

➢ Application in medicines:
- Genetically modified insulin:
Insulin synthesised as prohormone- contain A, B, C peptide
On maturation C peptide is removed
Challenge- removing C peptide in mature insulin using r-RNA
technique
Eli lily produced two RNA sequence corresponding to A and B chains
and introduced them in E. coli
Chains A and B -produced separately -combined by disulphide bonds-
human insulin
- Gene therapy:
Collection of method that allow correction of gene defect
Delivery of a normal gene into individual or embryo to take over the
function of and compensate for the non-functional gene.
Lymphocytes are grown in culture. A functional ADA cDNA (using
retroviral vector) is then introduced into these lymphocytes which are
subsequently returned to patients.
Patient requires periodic infusion of these as they not immortal.
If gene isolate from marrow cells producing ADA is introduced at early
embryonic stage, this could be permanent.
-Molecular diagnosis:
 methods of early diagnosis-
• recombinant DNA technology
• polymerase chain reaction
• ELISA- enzyme linked immuno- sorbent assay
very low concentration of bacteria and virus can be detected by
amplification of nucleic acid by PCR
PCR used to detect -
• HIV in suspected AIDS patients
• Mutation in genes of suspected cancer patients
A single stranded DNA or RNA tagged with radioactive molecule is
allowed to hybridise on its complimentary DNA in a clone of cells
followed by detection using autoradiography.
ELISA is based on the principle of antigen-antibody interaction
➢ Transgenic animals-
- Animals that have had their DNA manipulated to poses and
express on extra gene
- Examples- Transgenic rats, rabbits, pig, sheep, cow and
fish.
- Developed for
• Study normal physiology and development
• Study of disease
• Produce biological products
• Vaccine safety (transgenic milk are used to test the
safety of polio vaccine)
• Chemical safety testing

➢ Ethical issues-
- GMO have unpredictable results when they are in ecosystem
- GOI has setup organisations like GEAC
- GEAC take decision regarding validity of GM research and
safety of GMO for public services
➢ Biopiracy- use of bio-resources without proper authorisation
- American company got patent right on basmati
- Attempts of taking patent on neem and turmeric my foreign
companies
- Brazzein protein (west African plant) was patented in USA

Points to remember
➢ Green Revolution- tripled the food supply.
Plant, bacteria, fungi and animals- gene manipulation: Genetically modified
organisms (GMO)
etc.
 Some of the strains of Bt produce proteins that kill certain insects-
Lepidopterans (tobacco budworm, armyworm)
Coleopterans (beetles)
Dipterans (flies, mosquitoes
cry IAc and cry IIAb – cotton bollworm
cry IAb- cotton borer
Nematode Meloidegyne incognitia affect roots of tobacco
About 30 recombinant therapeutics -approved for human-use the. In
India,12 of these is presently being marketed.
ELISA- enzyme linked immuno- sorbent assay
Infection by pathogen can be detected by the presence of antigen (proteins
and glycoproteins) or by detecting the antibodies
95% of all existing transgenic animals are mice.
GEAC- Genetic engineering approval committee
There are an estimated 200,000 varieties of rice in India alone.
Basmati rice is distinct for its unique aroma and flavour and 27
documented varieties of Basmati are grown in India.
Transgenic models exist for many human diseases such as cancer,
cystic fibrosis, rheumatoid arthritis and Alzheimer’s.
Human protein (α-1-antitrypsin) used to treat emphysema.

YEAR WHY IS IT IMPORTANT?

1983 Eli Lilly, an American company prepared genetically engineered insulin

1990 First clinical gene therapy was given to a 4-year-old girl with adenosine
deaminase (ADA) deficiency.

1997 The first transgenic cow, Rosie, produced human protein-enriched

milk (2.4 grams per litre). The milk contained the human alpha-
lactalbumin and was nutritionally a more balanced product for human
babies than natural cow-milk.

1997 An American company got patent rights on Basmati rice through the US
Patent and Trademark Office. This allowed the company to sell a ‘new’
variety of Basmati, in the US and abroad.