BIOTECHNOLOGY & ITS

APPLICATIONS
 CRITICAL RESEARCH AREAS OF
BIOTECHNOLOGY
• THREE AREAS
• Providing best catalyst as improved organism ,usually a microbe or
pure enzyme.
• Creating optimal conditions by engineering for a catalyst to act.
• Downstream processing technologies to purify the protein / organic
compound.
BASIC APPLICATIONS OF BIOTECHNOLOGY
Biotechnology has tremendous scope in different fields.
A ] Genetically modified crop.
New trait
Purpose of GMO crops
Genetically modified organisms –for proteins,
biomolecules & research.
BIOREMEDIATION
Bioremediation use of naturally occurring or genetically
engineered micro organisms to degrade pollutants.
It is grouped in to in situ & ex situ ANAND CHAKRABARTY
SUPER BUG PSEUDOMONAS PUTIDA
• [ Alcanivorax borkumensis bacteria - Bioremediation of oil spills]

• Energy production-biofuels as alternative sources of fuels derived frm

biomass that is plant or animal waste.
MOLECULAR DIAGNOSIS

c] Diagnostics – like PCR &ELISA tests are some of the techniques

that are reliable and help in early diagnosis

PCR-Polymerase chain reaction is useful for the diagnosis of

inherited disorders,viral bacterial diseases etc

ELISA –Enzyme linked immunosorbent test

Very important diagnostic tool to test , a person is HIV positive or

negative.
Bio technology applications in agriculture
• Increase in food production can be brought by
A]Agro chemical based agriculture.
B]Organic agriculture.
C]Genetic engineered crop based agriculture.
 AGROCHEMICAL BASED AGRICULTURE

Made up of plant breeding techniques like hybridisation

along with use of agro –chemicals and better management
practices.
Agrochemicals are expensive
Harmful effect on environment,
Genetically modified crops
 GREEN REVOLUTION
• M.S.Swaminathan -Father of green revolution
Use of improved crop varieties by cross hybridisation
• Use of agrochemicals
• Better management practice
• Threefold increase in food supply
ORGANIC AGRICULTURE
• Aimed at reducing the need for chemical fertilisers and pesticides which are
not only harmful to mankind but also pollute the environment causing long
term effects.
• Organic agriculture though very useful is often expensive
• Needs a very good knowledge of crops and pest / pathogen management
GENETICALLY ENGINEERED CROP BASED
AGRICULTURE
• Is an alternative path of increasing the yield and reducing the
dependence and harmful effects of chemical fertilisers and pesticides.
• Modern techniques have 2 unique advantages
• 1]Any
. gene ,from an organism chemically synthesised can be inserted
in to another plant.
• 2]Plants with exact desired traits can be created.
GENETICALLY MODIFIED ORGANISMS /
TRANSGENIC ADVANTAGES
1]Tolerance to abiotic stress
• 2]Pest resistance crops
• 3]Reduce post harvest losses
• 4]Increased efficiency of mineral usage by plants
• 5]Enhanced nutritional value of food
• 6]Production of desired colour flowers
7]Production of pharmaceutically important compounds
• 8]production of antibodies & vaccine
• 9]To enable non –nitrogen fixing plants to fix nitrogen’
APPLICATIONS OF BIO TECHNOLGY IN
AGRICULTURE
• A] Genetically modified cotton [insect resistant]
• B]Pest resistant plants
Bacillus thuringiensis [Bt]
• Bt toxin is produced by soil bacterium called Bacillus thuringiensis.
Bt toxin gene has been cloned from the bacteria and has been
transferred to plants to provide resistance to insects,
without need for insecticides in effect created as bio pesticide.
Ex Bt cotton ,Btcorn, rice tomato potato etc..
Bt toxins are insect group specific
• Some strains of Bacillus thuringiensis produce proteins that kill
certain insects like lepidopterans [tobacco budworm,army worm]
• Coleopetrans[beetles]& dipterans[flies and mosquitoes]
 Action of Bt toxin protein on insects
• Bt forms a protein crystals which contain toxic insecticidal protein.
• Initially Bt toxins / Cry toxins are inactive prototoxins.
• After ingestion by insect their inactive toxin becomes active.
• Due to alkaline pH in gut ,which solubilises crystals.
• Activated toxins binds to the surface of the midgut epithelial cells,
thus creating pores which causes swelling and lysis further leading to
death of the insects
 TYPES OF Bt TOXIN GENES [crygens]
• Specific Bt toxins genes obtained from Bt are used in several crop
plants like cotton .
• Bt toxin is coded by a gene named cry the toxin also called cry
protein.
• The proteins encoded by the genes cryIAc and cryIIAb produce toxins
to control the cotton bollworms & cryIAbproduce toxins to control
corn borer.
 Effect of Bt cotton on bollworm,the pest of
cotton plant. MODE OF ACTION
• Moth lays eggs on leaves

• Larvae emerge when eggs hatch. larvae are known as boll worms

• Bt protein is ingested by bollworm feeding on plant

• Bt binds to the enzymes in the gut and disrupts inner lining

• Bollworm stops feeding immediately, dies within 48-72 hours of feeding

MELOIDEGYNE INCOGNITIA-NEMATODE
PARASITE
 PEST RESISTANT PLANTS BY
RNA INTERFERENCE
• Effect of a nematode parasite
• The nematode parasite Meloidegyne incognitia infects roots of
tobacco plants and reduces yield.
• It can be prevented by using RNA interference [RNAi] process which
is checked by silencing of specific mRNA due to a complementary
dsRNA
• dsRNA binds&prevents translation of mRNA.
• Source of this complementary RNA [dsRNA] could be from an
infection by viruses having RNA genomes or mobile genetic elements
[transposons] that replicate via an RNA intermediate.
When specific mRNA is silenced it will not produce the protein
/enzyme formed by it .
• Protein / enzyme is important for the functioning process, the
process will get stopped due to lack of protein.
 ROLE OF RNA interference to make plants
nematode resistant.
• It was done by using bacteria Agrobacterium as a vector to
introduce nematode specific gene in to host plant.
• The gene or DNA introduced produced both sense and anti-
sense RNA in the host cells.
• These two RNAs are complimentary to each other and
when present together form a double stranded RNA called
dsRNA.
dsRNA initiates RNA interference that is it silences specific mRNA of
nematode .

specific mRNA is essential for nematode ,the consequence is parasite

nematode cannot survive in the transgenic host expressing specific
interfering RNA .

Hence transgenic plants gets protected from the nematode Parasite.

 BIOTECHNOLOGICAL APPLICATIONS IN
MEDICINE
• The rDNA technology is used for production of therapeutic drugs which
are safe & effective.
• About 30 recombinant therapeutics have been approved for human use
in the world, in our country 12 are being marketed presently.
• Molecules Therapeutic applications
• Insulin Diabetes
• Erythropoietin Anaemia
• Hepatitis B vaccine Immunisation against Hepatitis B
• Human growth hormone Growth hormone deficiency in children
GENETICALLY ENGINEERED INSULIN
• Insulin used for diabetes was earlier extracted from pancreas of
slaughtered cattle and pigs.
• Was not as effective as secreted by human body.
• Insulin isolated from other animals elicited immune response in the
human body in some patients
• Caused allergy in some patients
• Hence there was a need to produce human insulin i.e produced by
human gene
STRUCTURE OF INSULIN

• Hormone Insulin consists of 2 short polypeptide chains, chain A &

chain B which are linked together by disulphide bridges.
In mammals ,the insulin is first synthesised as a pro hormone
[just like proenzyme],it contains an extra stretch called C peptide
 Pro-hormone needs to be processed before it becomes fully mature
& functional hormone.
For this C –peptide is removed during maturation of insulin .
Mature insulin doesn’t have C-peptide.
Challenge to produce mature human insulin
• The main challenge in production of human insulin by rDNA was
getting insulin assembled in to a mature form.
• For this in 1983,an American company Eli lily prepared two DNA
sequences corresponding to chain A &B of human insulin and
introduced them in plasmids of E.coli to produce insulin chains.
• Chains A&B were produced separately ,extracted and then combined
creating disulphide bonds to form human insulin .
.
Gene therapy
• For the first time in 1990, M.Blease & W.F.Andresco of National Institute of
Health, attempted gene therapy on a 4 year old girl
with ADA-[ADENOSINE DEAMINASE DEFICIENCY]
CAUSE OF ADA DEFICIENCY
Caused due to deletion of gene for ADA
This enzyme is involved in the maturation of T&B lymphocytes
Can be cured by bone marrow transplantation &enzyme replacement
therapy but it is not fully curative.
MAJOR EFFECT ON THE BODY
• Cannot produce B & T lymphocytes hence they have weak immune
system.
• Such children should live in germ free conditions of plastic bubble as
they cannot make ADA ,an enzyme essential for immune system.
Subjected to life threatening disease even if it is mild cold and
cough.
• Hence ideal approach is to give the patient a functioning ADA gene to
produce ADA in the body
STEPS IN GENE THERAPY
• In gene therapy ,lymphocytes from patients blood were grown in a culture
and functional ADA,cDNA was introduced in lymphocytes using a retrovirus
vector.
• The lymphocytes were transferred in to the patients body.
• Periodic infusion of such genetically engineered lymphocytes is done
because these cells are mortal.
• For permanent cure, gene isolated from bone marrow cells producing ADA
introduced in to cells ,at early embryonic stage can be a possible cure.
• Other disease like cancer, haemophilia Parkinson’s disease,
• cystic fibrosis etc ., are also treated by gene therapy.
TRANSGENIC ANIMALS
COMMON REASONS FOR DEVELOPING
TRANSGENIC ANIMALS
• STUDY OF NORMAL PHYSIOLOGY AND DEVELOPMENT
STUDY OF DISEASES
BIOLOGICAL PRODUCTS
VACCINE SAFETY
CHEMICAL SAFETY TESTING
NORMAL PHYSIOLOGY & DEVELOPMENT
• Transgenic animals can be specifically designed to allow the study of
• A]How genes are regulated ,
B]How they affect normal functions of the body & its development.
For e.g. Transgenic animals are used in study of complex growth
factors like insulin-like growth factor
 Study of diseases
• Many transgenic animals are designed to increase our understanding
of ,study of genes which are responsible for diseases in humans and
their treatment.

• Transgenic models exist for many diseases such as cancer, cystic

fibrosis, rheumatoid arthritis & Alzheimer’s disease.
BIOLOGICAL PRODUCTS
• Medicines required to treat certain diseases can contain biological
products ,often expensive to make.
• Useful biological products can be produced by introducing ,into
transgenic animals, the portion of gene [DNA] which codes for a
particular product.
• For e.g human protein [alpha-1-antitrypsin] is used to treat
emphysema.
 ROSIE
In 1997 ,the first transgenic cow ROSIE produced
human protein-enriched milk[2.4 gm per litre].
The milk contained the human alpha-lactalbumin
and was more nutritionally balanced for human babies
than natural cow milk
VACCINE SAFETY
• Transgenic mice are being developed for use in testing the safety of
vaccines before they are used to humans.
• e.g polio vaccine.
• If successful & found to be reliable they could replace the use of
monkeys to test.
CHEMICAL SAFETY TESTING
• Known as toxicity / safety testing
• Transgenic animals are made to carry genes which make them more
sensitive to toxic substance than non –transgenic animals.
• Exposed to toxic substance & effects studied.
• Toxicity testing in such animals will allow us to obtain results in less
time.
ETHICAL ISSUES
• Genetic modification of organism show unpredictable results ,when
such organisms are introduced in to ecosystem.
• Modification& use of living organisms for public services creates
problem with patents granted[food& medicine]
• GEAC [Genetic Engg Approval Committee] was formed by our govt to
decide the validity& safety of GMO for public safety.
• There is growing public anger that certain companies are being
granted patents for products & technologies that make use of the
genetic materials, plants & biological resources - which are grown,
identified & used by farmers & indigenous people related to a specific
region / country.
Basmati rice
Basmati rice is unique for its aroma & flavour
27 varieties are cultivated in India
• There is reference to basmati in ancient texts folklore & poetry as it
has been grown for centuries.
• In 1997 an American company got patent rights on Basmati rice
through the US patent & Trademark office and was allowed to sell a
new variety in US & abroad.
• This new variety of basmati was derived from Indian farmer’s
varieties.
• Indian basmati was crossed with semi-dwarf varieties & claimed as an
invention or a novelty
Besides Basmati rice now attempts are in progress for turmeric &
neem.
If we are not vigilant & we donot immediately counter these patent
applications ,other countries / individuals may encash on our rich
legacy & we may not be able to do anything about it.
NEEM
BIOPIRACY
• Financially rich nations are poor in biodiversity &traditional
knowledge.
• In contrast, Developing &Underdeveloped nations are rich in bio
diversity &traditional knowledge related to bio resources .
• Knowledge can be exploited
• To develop modern applications &can also be used to save time,
effort,expenditure during their commercialisation
• Some nations are developing laws to prevent such unauthorised
exploitation of their bio resources &traditional knowledge.
PATENT
• Set of exclusive rights granted by a state[national government] to an
inventor or their assignee for a limited period of time in exchange for
a public disclosure of an invention.
• Patents satisfy three criteria
• Novelty
• Non-obviousness
• Utility

• THANK YOU