Biotech Lecture Development

VISTAS
IN
BIOTECHNOLOGY
19th March, 2008

Biotechnology
Bio = Biology
Technology = Application
The application of Biology
(for the benefit of humans)
Biotechnology - a definition
The application
of
science & technology to living organisms
as well as
parts, products and models thereof,
to alter living or non-living materials
for
the production of knowledge, goods and
services
What is biotechnology?
Biotechnology is a multidisciplinarian in nature, involving
input from
Engineering
Computer Science
Cell and Molecular Biology
Microbiology
Genetics
Physiology
Biochemistry
Immunology
Virology
Recombinant DNA Technology Genetic manipulation of
bacteria, viruses, fungi, plants and animals, often for the
development of specific products
What are the stages of biotechnology?
Ancient Biotechnology
early history as related to food and shelter,
including domestication
Classical Biotechnology
built on ancient biotechnology
fermentation promoted food production
medicine
Modern Biotechnology
manipulates genetic information in organism
genetic engineering
Ancient biotechnology
History of domestication and agriculture
Paleolithic society Hunter-gatherers Nomadic

lifestyle due to migratory animals and edible plant
distribution (wild wheat and barley) (~2 x 106 yrs.)
Followed by domestication of plants and animals
(artificial selection) People settled, sedentary
lifestyles evolved (~10,000 yrs. ago)
Cultivation of wheat, barley and rye (seed
collections)
Sheep and goats milk, cheese, button and
meat
Grinding stones for food preparation
New technology Origins of Biotechnology
Agrarian Societies
Fermented foods and beverages
Long history of fermented foods since people

began to settle (9000 BC) (fervere to boil)
Often discovered by accident!
Improved flavor and texture
Deliberate contamination with bacteria or
fungi (molds)
Examples:
Bread
Yogurt
Sour cream
Cheese
Wine
Beer
Sauerkraut
Fermented foods and beverages
Dough not baked immediately would undergo

spontaneous fermentation would rise
Eureka!!
Uncooked fermented dough could be used to

ferment a new batch no longer reliant on
chance fermentation
1866 Louis Pasteur published his findings on

the direct link between yeast and sugars CO2 +
ethanol (anaerobic process)
1915 Production of bakers yeast

Saccharomyces cerevisiae
Classical biotechnology
Industry today exploits early discoveries of the fermentation
process for production of huge numbers of products
Different types of beer
Vinegar
Glycerol
Acetone
Butanol
Lactic acid
Citric acid
Antibiotics WWII (Bioreactor developed for large
scale production, e.g. penicilin made by fermentation
of penicillium)
Today many different antibiotics are produced by
microorganisms
Cephalosporins, bacitracin, neomycin,
tetracycline..)
Chemical transformations to produce therapeutic

products
Substrate + Microbial Enzyme Product
Examples:
Cholesterol Steroids (cortisone, estrogen,
progesterone) (hydroxylation reaction -OH
group added to cholesterol ring)
Microbial synthesis of other commercially valuable

products
Amino acids to improve food taste, quality or

preservation
Enzymes (cellulase, collagenase, diastase,

glucose isomerase, invertase, lipase, pectinase,
protease)
Vitamins
Pigments
Modern biotechnology
Cell biology
Structure, organization and reproduction
Biochemistry
Synthesis of organic compounds
Cell extracts for fermentation (enzymes
versus whole cells)
Genetics
Resurrection of Gregor Mendels findings 1866
1900s
Theory of Inheritance (ratios dependent on traits of
parents)
Theory of Transmission factors
W.H. Sutton 1902

Chromosomes = inheritance factors
T.H. Morgan Drosophila melanogaster

Molecular Biology
Beadle and Tatum (Neurospora crassa)

One gene, one enzyme hypothesis
Charles Yanofsky colinearity
between mutations in genes and amino
acid sequence (E. coli)
Genes determine structure of proteins
Hershey and Chase 1952

T2 bacteriophage 32P DNA, not 35S protein
is the material that encodes genetic
information
Watson, Crick, Franklin and Wilkins (1953)

X-ray crystallography
1962 Nobel Prize awarded to three men
Chargaff DNA base ratios
Structural model of DNA developed
DNA Revolution Promise and Controversy!!!
Scientific foundation of modern biotechnology

based on knowledge of DNA, its replication,
repair and use of enzymes to carry out in vitro
splicing DNA fragments
Biotechnology Timeline
1750 BC The Sumerians brew beer.
500 BC Chinese use moldy soybean
curds as an antibiotic to treat
boils
1590 Janssen invents the microscope
1675 Leeuwenhoek discovers cells
(bacteria, red blood cells)
1830 Proteins are discovered
1833 The first enzymes are isolated
1855 The Eschirium coli bacterium
is discovered
1859 Charles Darwin publishes On

the Origin of Species
1864 Louis Pasteur shows all living

things are produced by other
living things
1865 The age of genetics begins
1902 Walter Sutton coins the term

gene - proposed that
chromosomes carry genes
Gregor Johann
Mendel
1910 Chromosomal theory of
inheritance proved
1928 Fleming discovers antibiotic

properties of certain molds
1941 George Beadle and Edward Tatum propose

that one gene makes one protein
1949 Sickle cell anaemia demonstrated to be

molecular disease
1952 The Waring Blender
experiment
1953 The double helix is

unravelled
1967 The genetic code is cracked
1973 Recombinant DNA

technology begins
1975 First international conference

on recombinant DNA
technology
1975 DNA sequencing discovered
1975 Monoclonal antibody

technology introduced
1978 Genentech Inc. established
1978 Genentech use genetic engineering to produce

human insulin in E.coli - 1980 IPO of $89
1978 Kary Mullis discovers PCR

1996 First mammal cloned from adult

cells
1990s First conviction using genetic

fingerprinting
1996 Development of Affymetrix

GeneChip
1997 First artificial chromosome

Breaking the Genetic Code Finding the Central
Dogma
An RNA Club organized by George Gamow (1954)

assembled to determine the role of RNA in protein
synthesis
Vernon Ingrams research on sickle cell anemia (1956)

tied together inheritable diseases with protein structure
Link made between amino acids and DNA
Radioactive tagging experiments demonstrate

intermediate between DNA and protein = RNA
RNA movement tracked from nucleus to cytoplasm site of
protein synthesis
What are the areas of biotechnology?
Organismic biotechnology
uses intact organisms and does not alter genetic
material
Molecular Biotechnology
alters genetic makeup to achieve specific goals
Transgenic organism: an organism with artificially

altered genetic material
What are the benefits of biotechnology?
Medicine
human
veterinary
biopharming
Environment
Agriculture
Food products
Industry and manufacturing
What are the applications of biotechnology?
Production of new and improved crops/foods,

industrial chemicals, pharmaceuticals and livestock
Diagnostics for detecting genetic diseases
Gene therapy (e.g. ADA, CF)
Vaccine development (recombinant vaccines)
Environmental restoration
Protection of endangered species
Conservation biology
Bioremediation
Forensic applications
Food processing (cheese, beer)
Transfer of new Anti-cancer drugs
Culture of plants
genes into animal from single cells Diagnostics
organisms
Cell Monoclonal
Culture Antibodies
Crime solving
Molecular
Biology
DNA Tracers
technology Genetic
Engineering
Synthesis of
Banks of Cloning specific DNA
DNA, RNA Synthesis
probes
and proteins of new Mass prodn. of
proteins human proteins
Complete Localisation of
New types of Resource bank
map of the genetic disorders
plants and for rare human
human
animals chemicals
genome
New
New types antibiotics
of food Gene therapy
Red biotechnology is applied to medical processes. Some examples are the
designing of organisms to produce antibiotics, and the engineering of genetic
cures through genomic manipulation.
White biotechnology, also known as grey biotechnology, is biotechnology
applied to industrial processes. An example is the designing of an organism
to produce a useful chemical. White biotechnology tends to consume less in
resources than traditional processes used to produce industrial goods.
Green biotechnology is biotechnology applied to agricultural processes. An
example is the designing of transgenic plants to grow under specific
environmental conditions or in the presence (or absence) of certain
agricultural chemicals. One hope is that green biotechnology might produce
more environmentally friendly solutions than traditional industrial
agriculture. An example of this is the engineering of a plant to express a
pesticide, thereby eliminating the need for external application of pesticides.
An example of this would be Bt corn. Whether or not green biotechnology
products such as this are ultimately more environmentally friendly is a topic
of considerable debate.
Cloning
a collection of scientific
techniquesto create, improve, or
modify plants, animals, and
microorganisms
Cloning
Dolly the worlds first
cloned sheep at the
Roslin Institute
When?
1997
Where?
Scotland
Nuclear Transfer
Technique
involved fusing
a donated cell
with an egg
from which the
nucleus had
been removed.
How old was Dolly?
That depends on whether you mean her
birth age - six years - or her genetic age -
twelve years.
Dolly was born in 1997, but all the cells in
her body originate from genetic material
extracted from the udder of a six-year old
sheep.
One week earlier
Australia's first cloned ewe
dies mysteriously
Last Updated Fri, 07 Feb 2003
11:45:50 CANBERRA - Australia's
first cloned sheep has died
unexpectedly. Independent post-
mortem tests failed to identify the
cause of death, scientists said
Friday
Can Clones Reproduce?
The Australian ewe
bore 3 lambs
Dolly gave birth to 6
Here she is with her
first lamb, Bonnie
Cloning Endangered Species
On January 8, 2001,
scientists at Advanced
Cell Technology, Inc.,
announced the birth of the
first clone of an
endangered animal, a baby
bull gaur (a large wild ox
from India and southeast
Asia) named Noah.
Noah died of an infection
unrelated to the procedure.
Cloning Extinct Organisms
August 21, 2002
CNN.com reports
Japanese scientists are
planning to use tissue
from the legs and
testicles of a dead
mammoth to clone the
extinct creature and
display it at an Ice Age
wildlife park in Siberia.
COLLEGE STATION,
TEX. - Scientists in
Texas have cloned a
female calico kitten,
named "Cc:",
believed to be the
first pet successfully
cloned.
The kitten was born
on Dec. 22, 2001
The Missyplicity Project
http://www.missyplic
ity.com/M2.Pages/M2.
welcome.html
Korean scientist cloned a dog, but the
rest was fake, panel says
http://www.cbc.ca/story/world/national/2006/01/10/clone-fraud20060110.htm
l
Should we clone humans?
Who would we clone?
Cloning Isnt That Easy
Dolly was only
cloned after 276
tries
First Cloned Horse Created in
Italy Washington Post Staff Writer Thursday, August 7, 2003;
The clone with her

mother, whose skin
cell was used to make
the cloned embryo.
The two are twin
sisters.
Scientists Could Copy
Prizewinners
Cloning Yields Human-Rabbit
Hybrid Embryo
Washington Post Staff Writer
Thursday, August 14, 2003; Page A04
Scientists in China have, for the first time,
used cloning techniques to create hybrid
embryos that contain a mix of DNA from
both humans and rabbits
Hybrid Embryo
More than 100 of the hybrids, made by
fusing human skin cells with rabbit eggs,
were allowed to develop in laboratory
dishes for several days before the scientists
destroyed them to retrieve so-called
embryonic stem cells from their interiors
Hybrid Embryo
The vast majority of the DNA in the embryos is
human, with a small percentage of genetic
material -- called mitochondrial DNA --
contributed by the rabbit egg.
No one knows if such an embryo could develop
into a viable fetus, though some experiments with
other species suggest it would not.
Is cloning "unnatural?
Not at all - some organisms in nature only
reproduce using cloning - not only bacteria
and yeasts, but also larger organisms like
some snails and shrimp
A shrimp called Artemia perthenogenetica -
has survived for at least 30 million years.
Is cloning "unnatural?
In nature sexual reproduction is the only
way to improve the genetic stock of a
species, most asexual species tend to die off
Many more species, including the aphid,
reproduce by cloning most of the time, only
reproducing sexually every few generations
Sea cucumbers clone themselves: biologists
Now researchers have

determined that three
new groups of
echinoderms sea
cucumbers, sand
dollars and sea urchins
can spontaneously
clone themselves.
Gene gun
Transformation by Gene gun
INFORMATION TECHNOLOGY
MOLECULAR MEDICINE
NANOTECHNOLOGY
BIOMASS CONVERSION
Maize Crop
Grain Stover
Chemical
Pretreatment
Endosperm Germ
Containing Cellulose
Currently used suitable for
For ethanol production Cellulases
enzyme
degradation
Carbohydrates Fermentation Carbohydrates
ETHANOL
a n y sp e c ie s o f p la n ts are
M
er.
used for food and shelt
Yields and quality are
rt an t is su e s d e ali ng w ith
impo
plant biotechnology.
Important
crops inclu
de:
wheat, corn
,
rice, cotton
,
soybeans, a
nd
Nutraceutical provide benefits other than many kinds
of
nutrition ex disease fighting drugs vegetables
W H A T I S
BIOTECHNOLOGY?
A more precise
extension of traditional
plant breeding
Traditional plant
breeding randomly
transfers many
unknown genes to
enhance quality, size,
appearance
Biotechnology transfers
Traditional plant breeding
Traditional donor Commercial variety New variety
DNA is a strand of genes, (many genes are transferred)
much like a strand of
pearls. Traditional plant
breeding combines many
genes at once.
X =
Desired Gene (crosses)

Desired gene
Plant biotechnology
Desired gene Commercial variety New variety
Using plant biotechnology, (only desired gene is transferred)
a single gene may be
added to the strand.
=
(transfers)
Desired gene
Plant biotechnology definition
A precise process in
which scientific
techniques are used to
develop useful
and beneficial plants.
Plant Biotechnology:
1) Plant Tissue Culture
(Cloning)
2) DNA introduction
(Gene Transfer)
Cloning
in Plants
s?
gras
Ros
Turf
?es
Apple s?
s? fo di l
Orang D af
Have you ever
Potato es
eaten/used ?cloned plants?
es? rr i es ?
raw b e
St
Xmas Cactus Cotton Fern
Poplar Soybean Ohio Buckeye

Gene Identification:
Pro
t
ein
n ce Virus resistance
and
re sis t a
r b ic id e
He Gr
Oil
Value added ain
qu
stress re a l s ali
s i s ta n c e ut ic o r ty
c e e r c o l
rm a F lo w
e P ha
tr a t iv
Neu s
v ira l va c c in e
Genomics
im a l G r ow th h a bit
n
A
Insect Proteomics
resista
nce Metabolomics
Fu ng a l r e s is ta nc e
First biotech plant
product Flavr
Savr tomato
Herbicide Resistant Crops (HRC):
The concept
+ CP4 EPSPS = Roundup

gene Ready
Soybeans: Roundup Ready

Corn: Roundup Ready, Liberty Link
Cotton: BXN, Roundup Ready
Canola: Liberty Link, Roundup Ready
Interspecific Cross
Wheat Rye
Triticale
New species, but
NOT biotechnology
products
Agriculture Products On the Market
Insect resistant cotton

Bt toxin kills the cotton boll worm
toxin gene from a bacteria
Source: USDA
Insect resistant corn

Bt toxin kills the European corn borer
toxin gene from a bacteria
Rootworm GM approved (2/26/03)
Normal Transgenic
Herbicide resistant crops
current: soybean, corn, canola
coming: sugarbeet, lettuce, strawberry,
alfalfa, potato, wheat (on hold)
resistance gene from bacteria
Source: Monsanto
Virus resistance
papaya, squash, potato
resistance gene from a virus
Bacterial and Animal Biotechnology Products
Biotech chymosin
enzyme used to curdle milk products
gene from yeast
Source: Chr. Hansen harvested from GE bacteria
replaces the calf enzyme
bST (bovine somatotropin)

increases milk production
gene from cow
protein harvested from GE bacteria
Source: Rent Mother Nature
replaces cow protein originally
harvested from pituitary glands
of slaughtered cows
Next Generation of Ag Biotech Products
Golden Rice
Increased Vitamin A content
Transgenes from bacteria and daffidol
Controversory: large amount needed to
solve problem
Sunflower
White mold resistance
Resistance gene from wheat
Source: Minnesota
Microscopy Society
Turfgrass
Herbicide resistance
Slower growing
reduced mowing = reduced pollution
Bio Steel
Spider silk strongest known protein
Protein expressed in goat milk
Protein used to make soft-body,
bullet proof vests (Nexia)
ENVIRONMENTAL
HAZ ARDS
WILDLIFE WATER
ENDANGERMENT CONTAMINATION
SOIL EROSION INEFFICIENT

CHEMICAL USE
More food
Bt corn 3.5 billion pound yield
increase and $125 million in
additional income
Bt cotton 185 million pound yield
increase and $102 million in
additional income
Biotech soybeans $1 billion
in additional income through
production cost savings
Source: National Center for Food and Agricultural Policy
Products in the pipeline
Agronomic benefits
Oranges resistant
to citrus canker
Disease-resistant
sweet potatoes
Pest- and disease-
resistant cassava
Disease-resistant bananas
Benefits of biotechnology More food

Better food
Benefits of biotechnology Better food

Increasing production on existing
land preserves forests, enhancing
biodiversity
U.N. report says biodiversity
will be threatened on
72 percent of global land
area in 30 years.
HOW DOES IT WORK?
1.Identify the desired

beneficial trait
2.Isolate the specific
gene
3.Transfer the gene into
the cells of the second
plant.
4.Deliver new, improved
genetic traits to the
INCREASED USE
From one million acres to ninety million acres in just five years.
Soybeans Cotton
54% 61%
Corn 25%
ATTRIBUTES 1
Pest Resistant
Insects destroy over
$1 billion in crops
each year.
Bt crops produce a
protein that
eliminates targeted
pests.
Bt naturally destroy
their own insect
predators.
ATTRIBUTES 2
Herbicide
Tolerant
Herbicide tolerance allows growers to
utilize lower cost pesticides that are gentler
on the environment.
Crops are genetically improved to tolerate

certain herbicides so that weeds are
controlled and the crop thrives.
ATTRIBUTES 3
Disease
Resistant
Crop diseases reduces
yield and quality.
Biotech transfers new
disease fighting gene
into crop plants.
Crops are genetically
enhanced to naturally
resist disease.
Tools of Molecular
Biology
BASIC VECTOR
right T-DNA border
promoter
leader sequence
intron (optional)
targeting sequence (optional)

base vector
product gene
10381 bp
Col E1 replication origin potato proteinase inhibitor I terminator
cauliflower mosaic virus 35S promoter

streptothricin acetyltransferase
phosphinothricin N-acetyltransferase
streptomycin / spectinomycin nucleotydyltransferase cauliflower mosaic virus 35S terminator
left T-DNA border
Genetic Marker Analysis
In a way, humans have been genetically engineering foods and
animals for thousands of years
Selective Breeding:
Crossing two parents who are dominant for desirable traits
such as high crop yield, resistance to disease, long life,
sociableness (in animals like dogs and cats), or high milk
production (in cows, etc.).
May lead to low genetic variability

Genetic Engineering:
Moving the DNA of one organism to another organism, not
necessarily of the same species.
These mice have been inserted with a jellyfish gene

which allows them to fluoresce under special light
sources. Normal mice appear black under the same
light.
National Geographic article about glowing mice

Spawning and
Incubation
Impacts of hatchery fish
on wild populations
Coho Salmon
Novel gene
construct:
strong promoter
(sockeye MT) +
extra growth
hormone gene
(sockeye GH).
Growth hormone expressed in cold

waters, unlinked from temperature cue.
Hybridization between
black and white crappies
Genetic Relationships
Among Populations of
Yellow Perch
Genetic Engineering???
Mouse and Human Ear
Looking more like the handiwork of a mad
scientist, this mouse actually served as a host
for growing a human ear. Dr. Charles Vacanti of
the University of Massachusetts implanted a
scaffold, or mold, in the shape of a human ear
on the back of the mouse. The scaffold was
made of fine strands of a new kind of
biodegradable plastic and seeded with living
cells. The living cells adhered to the plastic fibers
as they grew and took on the shape of the mold.
In time the plastic mold dissolved, and the living
cells took over and generated new tissue; the
new tissue in this case being a three-
dimensional, contoured ear. Where does the
mouse come in? ... The mouse provided the
warmth and nourishment for the growing cells.
http://tlc.discovery.com/convergence/superhuman/photo/zoom_03.html
Plasmid: Vector (carrier); small circular DNA that
reproduces outside the main bacterial genome.
DNA
Delivery
Agrobacterium Particle gun

Introduction of
the gfp gene into
different target
tissues
Petunia petal - cells
Soybean seed whole seedling

(on right) Wheat callus - cells
GFP expression in
wheat seeds (left
seed, on left) and
roots (below)
GFP expression in soybean tissues
No gfp red
chlorophyll
fluorescence
GFP green
fluorescence
gif animation of GFP expression in
soybean tissue
Shows variability in expression pattern
standard illumination on left gfp illumination on right
Plant Recovery
Starting Material
Immature seeds
Germination
Soybean
Embryogenesis
Development Induction
Proliferation
Benefits of biotechnology
More food
Better food
Better for the environment

Land Mine Detection
Without this effort,

that is dangerous to our military,
children are maimed.
Land Mine Detection
How biotechnology helps

Patented transgene added to plants
When metal from mine is detected
Plant turns from green to red
Technology developed by Aresa Biodetection
Mine detected
How can we get better crops?
I N C R EAS E D U S E
From one million acres to more than ninety million acres
in six years.
Soybeans Cotton
54%* 61%*
*74% intended *71% intended
2002 2002
Corn 25%*
*32 % intended 2002
*= per cent of 2001 actual acres *= per cent of intended 2002 acres, USDA
PLANT TISSUES
GM potatoes (above) with coat protein gene and
non-GM (below) exposed to common potato virus.
Insect resistance
Bt corn
Insect resistance from
Bacillus thuringiensis
Non-toxic to humans
Target insect: corn borer
40% U.S. Corn crop Bt
Potential to reduce
insecticide use
Cloning: Can science go too far?
W
HUMANS
BE
NEXT?
Oct. 4, 2000 Today on MSN

Double muscling
Selection
This is the oldest method
of plant improvement.
You take seeds from the
best plants and plant them
the next year.
Your crops will gradually
improve, but not in a
systematic way.
Breeding
Hybridization
Allows best traits of two
parent strains to be
present in crop plant
Hybrid vigor increases
productivity of the plant.
In corn, requires
hand detasseling,
often done by
teenage work crews
Cloning
Cloning, by growing new plants
from cuttings, has been done
since the beginning of
agriculture.
Any time you take a part of a
plant other than the seed and use
it to produce a new plant, you
are cloning.
Modern tissue culture is cloning,
but so is planting grapes from a
cutting.
Apples , Potato, Grapes,
Orchids, Oranges, Bananas,
African Violets are grown this
way.
Grafting
Grafting is simply splicing two

plants together in such a way that
they survive.
Most commonly used in fruit
trees, a stem which produces
good fruit is grafted onto a trunk
which typically grows short and
strong.
Radiation Mutagenesis
Radiation:
(Photo of Bikini Explosion)
Chemical Mutagenesis
Same idea as radiation, and about as old;

use chemicals to induce mutations, then
study the mutated offspring for desirable
traits
Completely random, but has resulted in
commercial varieties of wheat, lettuce and
many species of ornamental flowers.
VEGETABLES FRUITS
Tomato, Potato, Eggplant Apple, Strawberry,
Lettuce, Celery, Cauliflower Walnut, Muskmelon,
Cabbage, Sugarbeet, Carrot, Papaya, Grape
Cucumbers, Sweetpotato,
Cassava
Transgenic Crops
for Food
CEREALS
EDIBLE OILS
Wheat, Rice
Mustard
Maize, Rye
Oilseed rape
Canola
LEGUMES
Sunflower
Soybean, Pigeon pea,
Chick pea
Agencies of Public sector promoting
Agricultural Biotechnology
Ministry of Science and Technology
Department of Biotechnology
Department of Science And Technology
Department of Scientific and Industrial Research
(Council for Scientific and Industrial Research)
Ministry of Agriculture
Department of Agricultural Research and Education
(Indian Council of Agricultural Research and Education)
Biotech Industry in India (2004-05)
Segment Revenues ($ million) Market Share (%)
Growth
2003-04 2004-05 2003-04 2004-05 (%)
BioPharma 625.45 811.36 79.19 75.24 29.72

BioServices 62.50 96.59 7.91 8.96 54.55
BioAgri 29.55 75.00 3.74 6.95 153.85
BioIndustrial 54.09 72.73 6.85 6.74 34.45
Bioinformatics 18.18 22.73 2.30 2.11 25.00
Total Industry 789.77 1078.41 100.00 100.00 36.55

Size
BioPharma corners three-fourth of Indian market ($811
million out of $1070 million)
Transgenic research in India (Public Sector)
AAU, Jorhat, Assam
Bose Institute, Kolkata
Central Institute for Cotton Research, Nagpur
Central Potato Research Institute, Shimla
Central Tobacco Research Institute, Rajahmundry
Centre for Cellular and Molecular Biology, Hyderabad
Central Rice Research Institute, Cuttack
Delhi University, South Campus, New Delhi
Directorate of Rice Research, Hyderabad
IARI, New Delhi
IARI sub-station, Shillong
Transgenic research in India (Public Sector)
International Centre for Genetic Engineering and
Biotechnology, New Delhi
International Crop Research Institute for Semi-arid
Tropics, Hyderabad
Indian Institute of Horticulture Research, Bangalore
Jawaharlal Nehru University, New Delhi
Madurai Kamraj University, Madurai
Narendra Dev University of Agriculture, Faizabad
National Botanical Research Institute, Lucknow
Punjab Agricultural University, Ludhiana
Tamil Nadu Agricultural University, Coimbatore
TERI, New Delhi
University of Agricultural Sciences, Bangalore
Transgenic research in India (Pvt. Sector)
Ankur Seeds Limited, Nagpur
Hybrid Rice International, Gurgaon
Indo American Hybrid Seeds, Bangalore
M/s MAHYCO, Mumbai
Metahelix Life Sciences, Bangalore
MAHYCO Research Foundation, Hyderabad
Monsanto, Mumbai
M/s Proagro PGS (India) Ltd., Gurgaon
Syngenta India, Limited, Pune
Sungro Seeds Ltd, New Delhi
Target traits
Disease resistance
Improving the quantity of the protein
Increasing vitamin content
Stress tolerance
Herbicide resistance
Delayed ripening
Edible vaccine
Transgenic crops approved for field trials in 2005
Brinjal
Mahyco, Mumbai cry1Ac
Sungro Seeds Ltd, New Delhi cry1Ac
IARI, New Delhi cry1F
Cabbage
Cauliflower
Corn
Monsanto, Mumbai Cry1Ab
Metahelix Life Sciences, Bangalore Modified Mu-element
(Turbo-Mu)
Gene Splicing
A specific, known gene and its promoters are
inserted into the crop plant
Because the sequence of the inserted gene is
known, it is possible to locate it and to determine
its insertion site in the genome.
Subject to intense regulatory scrutiny
Electron micrograph of Agrobacterium tumefaciens infecting

From Recombining DNA to
Recombinant DNA Technology
1973 Cohen & Boyer
The Gene Pool becomes a Gene Ocean
Any organism on earth is a source for genes for
use by breeders
Recombinant DNA Technology is one of the most
powerful tools ever invented.
Genomics/Gene Expression
Genomics is the science of looking at the whole
genome of a species. Corn, Wheat and Rice,
among others, are the targets of genomic research
Genomics plus improved knowledge of
controlling gene expression may mean that genes
already in our crop varieties can be turned on to
protect them from pests and other hazards
(drought, salt).
Tissue Culture
Plant breeders have learned
how to use plant hormones to
grow whole plants from one
or a few cells.
Plant cells growing in culture
tend to produce a lot of
mutations, which can include
beneficial mutations.
Tissue Culture
Embryo Rescue
Embryo rescue enables breeders to attempt
wide crosses between varieties that could not
be hybridized before
Crosses are performed between plants but the
resulting embryo aborts before a seed is
produced. In embryo rescue, the embryo is
removed before this happens and grown
outside the parent plant to produce a new plant.
In this instance, you're crossing two plants that
are not sexually compatible, that is, species
that would not usually be expected to interact
in nature.
Tissue Culture
Anther Culture
Immature pollen from a diploid
(double chromosome) plant is
cultured and grown into whole,
haploid plants.
The whole plants with selected
desirable traits are then
manipulated to double their
chromosomes, becoming
conventionally-breedable plants.
Tissue Culture
Somoclonal variation
Variations occur in plants
regenerated from cultured
cells or tissues.
Resistance to early blight
(caused by Alternaria solani)
was developed from protoplasts
of Russet Burbank potato.
Mechanisms include chromosomal
rearrangements, somatic crossing
over, sister chromatid exchange,
transposable elements and gene
amplification that occur during callus
formation.
Tissue Culture
Protoplast Fusion
Isolated protoplasts can be induced to fuse. Non-
specific fusion of protoplasts from the same or
different species can be induced using two
methods:
Use of chemical fusing agents. However, this may have
an effect on protoplast viability.
By electrical depolarization (electro fusion). There is
little effect on protoplast viability. Gives an efficient
high frequency of fusion.
Tissue Culture
Protoplast Fusion
Cell Fusion can be used to:
Produce fertile diploid somatic hybrids of sexually
incompatible species.
Produce heterozygous lines within a single species which
could normally only be propagated by vegetative means
e.g. potato and other root crops.
Transfer of limited parts of the genome from one species to
another
Make novel interspecific and intergenetic crosses between
plants impossible to hybridize conventionally.
p o r t a n t p ar t
An im
u a l i ty o f l ife
of q
s t he a c c e s s to
i
h e a lt h c a re.
good es
c h p r o m o t
(Whi
).
good health
be
Health can
h en
impaired w Quality
a s e s tr i k e s . of life in
dise healthca cl u
re practi des
prevent c es t h at
disease
treat and a s we l l
h ea l d i s as
ease.
Environmental Applications
Indicator bacteria
contamination is detected in the environment
microbes sensitive to certain pollutants
Bioremediation
cleanup contaminated sites
uses microbes designed to degrade
the pollutant
Air Quality Management Grow Fresh Air
ArecaPalm
(LivingRoomPlant) SansevieriaTrifasciata
(BedRoomPlant)
Using toxin removing plants to clean the air
Biotechnology originally developed by NASA, USA and research done by
PBC - STIP on plants suitable for the Delhi climate
142
Edible Vaccines A Biopharming Dream
Biotech Plants Serving Human Health Needs
A pathogen protein gene is cloned

Gene is inserted into the DNA of plant (potato, banana, tomato)
Plant must be isolated and highly regulated!
Humans eat the plant
The body produces antibodies against pathogen protein
Human are immunized against the pathogen
Examples:
Diarrhea
Hepatitis B
Measles
Future Health-related Biotech Products
Vaccines
Herpes
hepatitis C
AIDS
malaria
Tooth decay
Streptococcus mutans, the mouth bacteria
releases lactic acid that destroys enamel
engineered Streptococcus mutans
does not release lactic acid
destroys the tooth decay strain
of bacteria
e b e t w e e n birth
The tim The use of
ife
and death. L i s e n ov er biotechnol
h a s r
expectancy hich contribute
ogy has
s t y e a r s, w d to life
the pa la t i on. expectanc
e p o pu
adds to th are seekin
y. Scientis
ts
g n ew
products to
further
enhance b
iotechnolo
to extend l gy
ife
expectanc
y.
GENES CAN BE ISOLATED
FROM BACTERIA, VIRUSES,
FUNGI, PLANTS OR ANIMALS
AND MADE TO EXPRESS IN
CHOICE HOSTS
VARIOUS SEGMENTS INVOLVED IN
MARKETING LABORATORY RESEARCH
Science Managers (Based on needs Technical

Conceiving a project feasibility vizs--vis infrastructure and market
Identification of a trends which other institions are working to
1 produce it value addition / cost economics)
Product/Process
Laboratory Sound Scientific Plan (Molecular

2 Biology)
Research
Legal Protection/ Patent
Upscale; Chemical Engineers (Economy, bye-
3
Fermentation product, other products)
4 Formulation Ease of Distribution (Shelf

Life; persistence, mode of
application)
Field Trial Compliance with

5 approval/ Regulatory
Procedure
Packaging &
6
Marketing By the Nonbiologists,
Attractive/ aggressive
Golden Rice
Engineered to produce more vitamin A
precursor, beta-carotene
In Southeast Asia, 70% of children under the age
of five suffer from vitamin A deficiency
Vision impairment and increased disease
(diarrhea) are the result of deficiency
Variety will be released to subsistence
farmers, may correct deficiencies in diets
Potato leaves suspended in wall-dissolving enzyme s
Genomics leads are advancing
Target Screen
Yield Quality
Photosynthesis Starch / Carbohydrates
Seed development Lipids / Oils
Yield Qualit
Plant form Protein
y
Nutrient utilization
Harvest ability Stress
Pest Stress Heat tolerance
Pest Cold tolerance
Disease resistance Drought tolerance
Insect resistance Nutrient conversion
Control + Gene Control

Control ++Gene
Gene
A CC
Photosynthesis Nutrient Utilization Environmental Stress
(More Biomass) (More Root Hairs) (Drought Stress)
Biotechnology:
The Invisible
Revolution
Corn is corn
Products
Soya is are
soyaproducts whatever the
BioPlastic is plastic
productionBioDiesel
technology.
is diesel.
Medicines are medicines

Biotech is delivering on its
Promise
Biotechnology has delivered significant
environmental benefits:
- insecticide reduction
- soil conservation and enhanced
productivity
- improved water quality
- enhanced sustainability
- promotion of reduced risk herbicides and
insecticides- improved feed quality
Many of these benefits are consistent with

Biotechs
primary goals
Biotech Crops have significantly improved

farm
competitiveness
Poultry includes
chickens, ducks, and
Animal
t e c h n o l ogy turkeys. These species
bio
e n u se d to produce meat, eggs,
has be
and feathers.
improve
s t o c k a nd
liv e
poultry.
Livestock
includes be
ef
cattle, dair
y
cattle, hog
s
and sheep
Vaccines protect
people from
specific diseases.
Biotechnology
Different diseases
provides new
need different
methods for
vaccinations. These
making vaccines
vaccines contain a
and leads to the
dead or less
development of
active copy of the
new vaccines
virus.
against different
diseases.
Antibiotics
Antibiotics and vaccines are among the biggest

medical advances since 1000. (Culver Pictures)
BC Yang
Thanks to work by Alexander Fleming (1881-
1955), Howard Florey ( 1898-1968) and Ernst
Chain (1906-1979), penicillin was first produced
on a large scale for human use in 1943. At this
time, the development of a pill that could reliably
kill bacteria was a remarkable development and
many lives were saved during World War II
because this medication was available.
A. Fleming E. Chain H. Florey

A tale by A. Fleming
He took a sample of the
mold from the
contaminated plate. He
found that it was from the
penicillium family, later
specified as Penicillium
notatum. Fleming
presented his findings in
1929, but they raised little
interest. He published a
report on penicillin and
its potential uses in the
British Journal of
Experimental Pathology.
u t i c a l s w e re
Pharmace l a n t s or Biotechnolo
o m p
isolated fr p l e of techniques a
gy
e x a m re
fungi. An utilized to
e c hn o l o g y
biot manufacture
engineered l s is vitamin C a
a c e u t i c a nd
pharm vitamin B-2
m a n i n s u l in . .
hu a ns
a l l y h u m
Origin a t was
s u l i n t h
used in
from pigs.
Designer drugs, gene therapy
n o t a l w a ys
People die n s p l a n t s are
because Organ tra e r bec a u s e t h e re
t a n s w
organs, such t he be s av a i l ab l e to
fe w er o rgans d o u r
as liver, heart, are e e d t he m a n
w h o n
or pancreas people a lw a y s a c c e pt
e s d o n o t
fail to work bodi
w o r ga n s .
properly. ne
These lives
can be saved
by organ
transplants.
Materials play
an important
role in human
life. Some
examples of
materials are:
plastic, A polyes
ter polym
kind of p er
synthetic olymer u is the
make pla sed to
fabrics, silicon stics. So
example me
chips and s a re : d i a
waste ba pers, yar
computers. gs, elect d
and deca rical tape
c a n h e lp ls. ,
lo g y
Biotechno rials
d uc e m at e
pro es t e rs i n a
pol y
including fe ctive
Polymer product of a chemical
ost e f union of tow molecules to the
more c
manner. same compound or produce
another compound.
Array Technologies
Monitors the interaction of molecules in a

group, with specific molecular probes
Most advanced version: DNA array
measures the intensity of the mRNA
gene
It is an arrangement of DNA
molecules of a known sequence
Human Genome Project
Project to sequence
the entire human
genome (3.1 billion
base pairs)
Working Draft
Completed Opened many new doors for
in 2000 scientists
Diagnostic medicine
Genetics
Therapeutics
Methods of Arrays
In situ synthesized DNA probes
With and without photolithographic masks
Pre-synthesized DNA probes

Brown Method
Filter-based DNA arrays
www.dsm.com/en_US/html/dfs/genomics_dna_arrays.htm
In Situ (Affymetrix)-Building the
DNA Probes
www.nearingzero.net/sbunch5.html
http://services.ifom-ieo-campus.it/
In Situ (Affymetrix)
Then the wafer is flooded with a solution containing the
DNA of interest and the strands hybridize and create
covalent bonds with any strand that is complementary.
In Situ Synthesized Arrays
Was first commercialized by Affymetrix Inc.

(Santa Clara, CA)
Other similar methods

Ink-jet printing on the DNA chip without
photolithography
digital lighting processors (DLPs) shine light at
certain places on chip to direct nucleotides
Pre-Synthesized Arrays
Developed by Ed Southern
Used at Patrick O. Brown Laboratory and

Stanford University
Less expensive equipment

Pre-synthesized DNA probes (100-500 bases
long) attached to glass slides
No robotics involved
Improvements in Medicine
Diagnostics
Faster and more accurate diagnosis
Positive prognosis
Predictable symptoms
Ability to make arrangements before
things happen
Applications
Forensics
Human and animal diagnostics
Individualized medicines
Sequencing
single nucleotide polymorphism (SNPs)
(SNPs
cytogenetics
Microarray Future
Do metastatic cancer cells differ
genetically from original growth
cancer cells?
An indicative list of Biotechnologies
DNA (the coding): genomics, pharmaco-genetics, gene probes, DNA
sequencing/synthesis/amplification, genetic engineering.
Proteins and molecules (the functional blocks): protein/peptide
sequencing/synthesis, lipid/protein glyco-engineering, proteomics,
hormones, and growth factors, cell receptors/ signalling/ pheromones.
Cell and tissue culture and engineering: cell/tissue culture, tissue
engineering, hybridisation, cellular fusion, vaccine/ immune stimulants,
embryo manipulation.
Process biotechnologies: Bioreactors, fermentation, bioprocessing,
bioleaching, bio-pulping, bio-bleaching, biodesulphurization,
bioremediation, and biofiltration.
Sub-cellular organisms: gene therapy, viral vectors.
Bioengineering: biomaterials, biosensors, biomedical devices and
bioinstrumentaion.
Biofertilisers and biopesticides.
Amazing Biotechnology
more than 300 biotech drug products and vaccines currently in clinical trials
targeting more than 200 diseases, including various cancers, Alzheimers disease,
heart disease, diabetes, multiple sclerosis, AIDS and arthritis
responsible for hundreds of medical diagnostic tests that keep the blood supply safe
from the AIDS virus and detect other conditions early enough to be successfully treated
e.g. home pregnancy test
biotechnology foods such as papaya, soybeans and corn quite popular
Biopesticides and other agricultural products also are being used to improve our food
supply and to reduce our dependence on conventional chemical pesticides
clean up hazardous waste more efficiently by harnessing pollution-eating microbes
without the use of caustic chemicals
cleaner processes that produce less waste and use less energy and water in such
industrial sectors as chemicals, pulp and paper, textiles, food, energy, and metals and
minerals
DNA fingerprinting, a biotech process, has dramatically improved criminal
investigation and forensic medicine, as well as afforded significant advances in
anthropology and wildlife management
Global business US $ 64 in 2004
Biotechnology in India
Initiative in early 1980s
Agencies - DST, CSIR, ICAR, ICMR, UGC
National Biotechnology Development Board
1982
Department of Biotechnology 1986
Strategy :
Creation of Infrastructure
Human Resources Development
Promotion of R & D
Technology Transfer
Promotion of Industry
Public Private Partnership
Regulation
International Co-operation
Institutions & Manpower
Number of hospitals : 16,000
Number of doctors : 500,000
Number of research institutions : 200
Number of Scientists : 25,000
Medical graduates produced : 17,500 per
yr
Science graduates produced : 500,000
per yr
Number of Agriculture Univs. : 40
HRD Programmes
Post Graduate Teaching Programmes Ph D
Programme
Post Doctoral Fellowship
Short Term Training Programes
Industrial Training
Seminar, Symposium, Conference
Awards and Scholarships
Travel Support
Post Graduate Teaching
General Biotechnology 43
Agricultural Biotechnology 10
Animal Biotechnology 3
Medical Biotechnology 3
Marine Biotechnology 2
Neurosciences 3
Industrial Biotechnology 1
Biochemical Engineering 7
Pharmaceutical Biotechnology 1
Genetic Engg. & Bioprocess Dev. 1
Molecular & Bioprocess Tech. 1
Clinical Biochem. & Biotech. 1
IPR 1
TOTAL 77
65 Centres under the
Bioinformatics
Information System
Network (BTISnet)
12 Nodes on the Virtual
Private Network (VPN)
BIOGRID
77 Universities and
Institutions where post
graduate teaching in
biotechnology is
supported by DBT
Bioinformatics Network
Year of Start 1986
Total No. of Distributed Bioinformatics Centers- 65 with
each one is having focus on a specific area of biotechnology
One year Advanced diploma course on Bioinformatics - 5
Interactive Graphics facility 6
Mirror Sites 5
Super Computing facility - 1
BIOGRID INDIA 12 Nodes
Infrastructure
DBT has established 63 major infrastructure facilities
- 20 in plant Sciences
- 11 in medical sciences
- 32 useful for both areas and others
6 Culture collections-
- Blue green algae
- Marine cyanobacteria
- Agriculturally useful organisms
- Industrial microbes
- Filaria
- Tuberculii
2 Animal House Facilities

Other Major Facilities
17 Genetic counseling centers established benefiting 18000
families and 4500 tribal families
2 Microarray Facilities
5 Automatic DNA sequencing Facilities
6 genomic /proteomic facilities
10 facilities for drug and molecular design
8 Gene Banks for crops, medicinal and aromatic plants
3 High/medium throughput facilities for screening extracts
1 fMRI Facility
5 Centers of Plant Molecular Biology
4 Centers for Genetic Engineering
Research & Development
1986 - 2005
More than 2500 R&D projects launched in 20 identified
areas.
National Bioresource Development Board set up for
bioprospecting and bioresource utilization.
Establishment of gene banks for conservation and
inventorisation of bioresource.
Rice genome sequencing as a member of International
Initiative - sequencing of part of chromosome 11
completed.
Participating in International Consortium on Silk Worm
Genomics and Tomato Genome Sequencing.
Complete sequencing of Indian isolate of hepatitis C done.
Agriculture Biotechnology - Crops
Focus :
- Improved Productivity
- Drought, Salinity Tolerance
- Reduced Yield due to Pests/Diseases
- Nutritional Enhancement
Crops :
- Rice, Wheat, Cotton, Mustard , Chickpea, Potato
Pigeonpea, Mungbean, Sugarcane
Strategy :
- Transgenics & Marker based Molecular Breeding
- Cloning and Characterization of New Genes and
Promoters
Crops
Wheat Rust: Durable resistance to leaf and stripe rusts using molecular
marker technology in bread wheat (DWR,DWR-RS, PAU, ARI &
NCL)
Rice: Salinity and dehydration stress tolerance: Cloning of responsive
genes, their promoters and development of transgenics (ICGEB,
UDSC, IARI, BI, UOH)
Sugarcane: 12,956 ESTs generated and deposited in genbank. 9000
genomic clones and several thousand cDNA clones developed from
high sugar and red-rot resistant lines
Crop Improvement for Better Nutrition in rice, sweet potato and
cassava using AmA1 Gene
Use of molecular marker technology approach in wheat quality
breeding
Transgenic Research in India
Target Crops/ Vegetables
Cotton, Corn, Mustard, Rice, Soybean, Potato, Tobacco, Coffee,
Tomato, Brinjal, Cauliflower, Pea, Cabbage, Banana,
Muskmelon, Pigeonpea, Chickpea, Bell-pepper, Blackgram,
Chilli, Watermelon etc.
Transgenes Employed
Bt. toxin genes, Herbicide tolerant genes (CP4 EPSPS, Bar
gene), Xa21, ctx-B and tcp of V.cholera, Chitinase, Glucanase,
ACC synthase, RIP, Protease Inhibitor, Lectin, Ama-1, OXDC
gene, Rabies glycoprotein gene, Bar, Barnase, Barstar, GNA
gene, Vip-3 gene, Bacterial Blight Resistance gene, Osmotin etc.
Transgenic Crops under Trial
Under green house trial : 19
Under limited field trial : 3
(Rice, Mungbean, Mustard)
Large scale field trial : 1
(Cotton)
Released for commercial production : 1
(Cotton)
Plant Biotechnology
Areas : Forestry, Horticulture & Plantation Crops
Status
57 regeneration systems documented
Protocols standardized for 20 plant species, 10 technologies
transferred to industry Eucalyptus, Bamboo, Teak,
Sugarcane, Potato, Black pepper, Coffee, Citrus
Ongoing R&D - Transgenics
Tomato, Grapes, Banana - shelf life
Chilli, Pepper, Ginger, Cardamom Disease resistance
Populus, leucaena - reduced lignin
Germplasm characterization Teak, Eucalyptus, Casurina
International Solanaceae Genome initiative launched
India to sequence chromosome 5 of tomato
Functional genomics of tomato nutritional quality, shelf life,
disease resistance
Production and demonstration of quality planting material of
Bamboo
Tomato Genome Sequencing
Animal Biotechnology
Diagnostics for Peste-des-Petitis Ruminants
(PPR) detection commercialized
Reconstituted Bovine collagen for wound and
burn healing commercialized
Multicentric programme on buffalo genomics
launched : Characterisation of traits of economic
importance
Multicentric programme on bovine tuberculosis
Programme on animal nutrition
Aquaculture & Marine
Bacterial immunostimulant for shrimp health management developed
Bioreactor system developed for water quality management in prawn hatchery
New source of insulin from carp adipose tissue
Quality agar extracted from seaweed (Gelidiella acerosa)
Efficacy of DNA-based vaccine for Aeromonas infection in carps under evaluation
Shrimp Genomics launched Comparative and functional genomics
Diagnostics and vaccines transferred to industry
Immunodiagnostic kit for detection of bacterial pathogens in finfish and
shellfish
Combi kit for simultaneous detection of White Spot Shrimp Virus and
Monodon Baculo Virus
Heat killed whole cell vibrio vaccine, shows immune response in shrimp
Immunodot detection kit for white spot shrimp virus
Technology ready for transfer
Bioreactor technology in shrimp hatcheries through bacterial conservation for
denitrification
Seribiotechnology
Improved quality and productivity in silk
Silkworm genome sequencing
Over 400 molecular markers generated
India participating in International Consortium on Lepidopteran
Genomics: More than 10,000 ESTs of muga silkworm and mori silkworm
characterized
Three high yielding hybrids of silkworm released to farmers
Network projects initiated for use of molecular markers in breeding of
disease-resistant silkworm and pest- and disease-resistant mulberry
Molecular characterization of non-mulberry silkworm (Tasar, muga,
eri)
Spider silk
Biophysical properties of spider silk proteins produced by Indian
species
Sourcing of spider silk genes and expression in Bombyx mori
Prospecting of Indian spiders venom for therapeutic purposes
Application of nanobiotechnology to spider silk
Medicinal & Aromatic Plants
Four national gene banks established - about 3500 accessions
Two specialized germplasm banks set up for plants used in Indian System of
Medicines
Production of high value therapeutic compounds in cell culture
(Podophyllotoxin, guggulsterones Z&E, camptothecin and azadirachtin)
Isolation and characterization of new thereapeutic leads (for anticancer,
antiamoebic, antidiabetic, bioenhancers, immunomodulatory).
Clinical trial of standardized herbal preparations - Terminalia arjuna left
ventricular dysfunction
Five technologies transferred to industry; several other leads under
discussion with industry
Herbals for veterinary health care - priorities identified
Functional genomics of selected medicinal and aromatic plants
Plants as bioreactor for production of biomolecules (antigens for cholera,
rabies), antibodies for therapeutics and industrial enzymes
Biofertilizers
23,000 ha demonstrated for use of biofertilizers;
50,000 farmers benefited
10 packages for Integrated Nutrient
Management developed for specific cropping
system of varied agroclimatic zones
A network project on development of efficient
strains of biofertilisers launched at 11 centres
Liquid biofertilizer technology
being developed
Biopesticides
Technology Development and Demonstration
24 Integrated Pest Management technology packages
developed; 7 transferred to industry
Over 150,000 ha covered in different agroclimatic zones;
65,000 farmers benefited
Insect sex pheromones successfully synthesized and
field-tested for the mass trapping of brinjal shoot and
fruit borer
More than 615 plant extracts prospected as source of
pesticides; 11 promising leads short listed for further
investigation
Product Development
Bioprahar: biopesticide (for cabbage and cauliflower)
developed by ICGEB, launched commercially (2004)
Bioprospecting
Bioprospecting of genes / molecules
96 stress related genes identified, characterized and
cloned, salt resistance gene transferred to rice and mung
bean, field trials of transgenic rice being conducted
Product development
Microbial biofertilizers for coastal region
Biopesticide for bollworm (Helicoverpa)
SOD antioxidant skin cream
Over 4000 plant species from Western Himalayas and
Western ghats screened for natural dyes; nine short listed as
potential sources
Network project on lac biotechnology initiated
Chemical and genetic profiling, conservation
6 endangered medicinal plant species
Microbial Diversity
Arid Zones : 48 Saline tolerant algal species, screened for pigment production, soil
conditioning and nitrogen fixation
Cold Habitats : > 100 bacterial species isolated from soil and water samples from
Antarctica
Hot Springs and sulfur springs : > 150 Actinomycetes species isolated, potential
rifamycin producer identified
Hydrocarbon contaminated Sites : 12 oil refineries screened, > 350 bacteria
species isolated, thermophilic Bacterial consortium developed for enhancing Oil
recovery from Oil wells
Marine Sources: > 500 Bacterial isolates screened for secondary metabolites,
benzoate and phenol degraders identified
Forest Belts: Virgin ecosystems, novel entomopathogenic and keratinolytic fungi
Isolated and identified
Enhancement of Oil Recovery from dead Oil wells - Bacterial Consortium
developed which on being injected in sick oil wells under high pressure leads to oil
recovery, tested in 25 Oil Wells of ONGC, helped in extraction of 4500 cubic meters
of oil worth US$675000
Environment Biotechnology
Conservation of endangered animals especially big
cats through embryo transfer technology and tissue
banking
Use of lichens as indicators of environmental pollution
Bioremediation of degraded ecosystems
Green technologies to treat industrial effluents (paper
& pulp, dyes, distilleries, tanneries etc)
Bioscrubber for removal of obnoxious industrial
emissions
14 technology packages developed; 5 under use by
industry
Programme on microbial approaches to combat global
warming
Medical Biotechnology
Vaccines
Diagnostics
Therapeutics
Stem Cell Technology
Biomedical Engineering
Nanomedicines
Clinical Trials and Clinical Research
Vaccines
Rabies : DNA vaccine, phase III animal
trials completed.
Cholera : Recombinant oral vaccine, Phase
II trial.
Malaria : Phase I clinical trials initiated.
Rotavirus : Phase I clinical trials underway
JEV : Vaccine technology transferred to
industry
HIV/AIDS : DNA candidate for subtype C
developed
Anthrax : Phase III clinical trials initiated.
Infectious Bovine Rhinotracheitis :

Commercialised
Diagnostics
Transferred to the Industries & Under Development &
Launched in the Market : Negotiations
Western Blot for HIV-I & II : HIV/AIDS for Transfer :
Naked Eye Agglutination : HIV/AIDS DAT : Toxoplasmosis
System (NEVA) Haemagglutination Assay :
ELISA : Hepatitis C Leishmaniasis
ELISA : Leishmaniasis IFA : Rabies
ELISA : Alpha-feto Protein Species Specific Tests : Snake Bites
(Pregnant Women) PCR Assay : Tuberculosis
Multiplex PCR : STD
Transferred, yet to be launched : PCR : Leptospirosis
IgM Mac ELISA : Dengue In-vitro Assay : Anti-Inflammatory
Japanese Encephalitis Drugs
West Nile Immunochromatogrpahy/ :
IgM ELISA : Hepatitis A Tuberculosis
Urine Based ELISA : Reproductive / ELISA Assay
Hormones
PCR Assay : Typhoid
Diagnostics : New initiatives
Future Plan:
New Innovative Technologies, easy to perform rapid, affordable
including those that detect Multiple Diseases through Syndrome
Approach useful for Medical Practices and Public Health Programmes
New platforms :
Optical Thin Film Biosensors
Micro Electro Mechanical Systems (MEMS) for Disease Diagnosis
Quantum Dots
Single diagnostics
Tuberculosis, malaria, TORCH, Viral load in HIV,
Neonatal sepsis, Thyroid function, STD, Typhoid, Mycosis,
Cardiac screen, diabetic screen
Syndromic Detection:
Malaria, Typhoid, Dengue and Tuberculosis
HIV/HCV, HBV, Malaria and Syphilis infections in blood for
transfusion
Indigenous production of control panels, sera.
Stem Cell Research
To establish stem cell banking facility in the
country.
To develop well characterized stem cell
lines.
Potential use of embryonic stem cell lines to
generate various organs.
Regeneration of tissues for stem cells
obtained from aborted foetus
Basic understanding of the signaling
mechanisms and differentiation of stem
cells.
Bioengineering
Tissue Engineering and Stem Cells
Biomaterials for Therapeutics
Biomedical Devices
Biomedical Instrumentation
Biosensors
Nanobiotechnology
Nano-particles for diagnostic/therapeutic

use
Biosynthesis of Nano-particles
Biological templates for nano-particle
assembly
Bio-nano composites
Imaging/sensing of Nano-particles/bio-
molecules
Technology transfer
Products in Market resulting in import
substitution and value addition
HIV diagnostic kits-Western Blot and ELISA test ,
Liposome mediated Amphotericin B drug delivery
system,
Leprosy vaccine (1st of its kind in the world),
14 diagnostic kits for detection of Pregnancy and
contraceptive problems,
12 Packages for bio-remediation of petroleum oil spills,
6 bio-fertilizers,
5 bio-pesticides formulations
6 Plant Tissue Culture protocols
Biotech Products in Indian market
r DNA Hep B vaccine

Recombinant streptokinase
Erythropoietin
, , interferon
Haemophilus influenzae B vaccine
Human insulin
Human Growth Hormone
Human Interleukin
Streptokinase
Granulocyte Colony Stimulating Factor
Follicle Stimulating Hormone
Tissue Plasminogen Activator
Biosafety Regulations
Recombinant DNA Safety Guidelines (1990)
Revised Guidelines for Safety in
Biotechnology -toxicity and allergenicity
evaluation (1994)
Revised Guidelines for Research in
Transgenic Plants & Guidelines for Preclinical
and clinical data for r-DNA based vaccines,
diagnostics and other biologicals (1998)
Ethical policy on Human Genome and
Genetic research and services (2000)
Regulations being simplified in an ongoing
dynamic process.
Other initiatives
Establishment of patent facilitation cell to promote national
and international patents of indigenous innovations
112 patents filled - 7 international patents and 7 national
patents granted.
Revision of biosafety guidelines for transgenic plants.
Guidelines for clinical trials of recombinant DNA vaccines
formulated and published.
Single window application processing mechanism for
recombinant products setup.
Accession to Budapest Treaty on microorganisms and
establishment of International Depository Authority.
National bioethics committee setup : Ethical policies on
human genome, genetic research and services published.
Indian Biotechnology Industry
Number of biotech companies : 300
Global ranking in terms biotech companies : 11
Investment growth in last 5 years : 50 % per annum
Annual Turnover (Apr 04 Mar 05) : US $ 1.07 Billion
Annual increase : 37 %
Percentage of global business (2004-05) : 1.6 %
(2003-04) : 1.5 %
(2002-03) : 1.2 %
Exports : 43 %
Major areas : Biopharma, Bioservices, Bioindustries,
Bioagriculture, bioinformatics, Biosupplies
Indian Biotech Industry 2004-05
Indian Biotech Industry
Indian Biotech Grows
Areas of oppurtunity in India
Vaccines, dignostics and therapeutics
Biomedical devices, instruments and sensors
Bioartificial organs using tissue engineering and
stem cells
Clinical trial and contract research
Agriculuture disease and pest resistant, drought
and salinity tolerant seeds and plants
Biofertilisers and Biopesticides
Bioinformatics
International Collaboration
Europe : U.K., Germany, Switzerland, Sweden, Norway,
Poland, France, Russia, Belarus, Denmark
North America : USA
South America : Brazil, Argentina, Venezuela, Cuba
Asia:
Multilateral : ASEAN, SAARC, BINASIA,
Asian Cooperation dialogue (ACD)
Bilateral : Myanmar, Vietnam, Malaysia, Thailand,
Singapore, Mangolia, China, Japan, Sri Lanka,
South Korea, Syria, Israel, Iran
Africa : Sudan, Egypt, Tunisia, Mauritius
Australia : Australia
Some New Initiatives
National Biotechnology Development Strategy
A Policy Document and 10 year Road Map
A Central pool of high caliber Scientists
Small Business Innovation Research Initiative
(SBIRI) A funding mechanism for
supporting early stage Pre-proof-of-concept
research and product development in small
and medium private enterprises
Genetic engineering
takes mankind into

realms that belong
to God and God
alone
- Prince Charles
The Threat
that Biotechnology Is
The threat is not
represented by those in
favor of GM crops.
Nor is it represented by those
opposed to GM crops.
The threat is a mutual creation
of both sides, namely, that
through our rhetoric, we may
erode the foundation of mutual
trust apart from which
democratic institutions fail.
If the unaccredited public becomes
involved in debate on matters as close to
the boundary between science and trans-
science as the direction of biological
research, is there some danger that the
integrity of the Republic of Science will
be eroded?
- UK and US scientific leaders at a meeting at the
CIBA Foundation, London, 1971
This is a grave threat to the integrity, balance, and
future evolution of life on earth...
The most significant risks are

-- Disruption of the ecosystem;
-- Genetic pollution;
-- Increased use of toxic, carcinogenic, and
mutagenic agricultural chemicals,
leading to water pollution . . .
-- Increased incidence of cancer, birth
defects, and other illnesses.
- John Fagan, Genetic Engineering: The Hazards;

Vedic Engineering:The Solutions (1995)
The threat is the all-or-nothing
rhetorical trap which both
sides have collaboratively
created, however unwittingly.
The Precautionary Response
When faced with two contrasting opinions about
issues related to food safety, consumers place great
emphasis on negative information.
They do so even if the source of the negative

information is known to be unscientific.
This effect appears to be particularly strong when a

consumer sees little to gain from a new food
technology.
When a given food is plentiful, it is rational to
place extra weight on negative information about
any particular piece of that food.
It is rational to do so even if the source of the

negative information is known to be biased.
The reasons are: I have nothing to gain from
eating an allegedly tainted piece of food.
And there is no way to prove the tainter wrong.
We cannot prove the tainter wrong in spite of the
factindeed, because of the factthat the tainter
offers no evidence.
This suggests that a single person with a negative
view will be much more influential than many
people with a positive view.
Example of precautionary response:
Irradiated pork study
The neutral description
The U. S. FDA has recently approved the
use of ionizing radiation to control
Trichinella in pork products. Results is
10,000 fold reduction in Trichinella
organisms in meat. Process does not induce
measurable radioactivity in food.
The positive description

Each year, 9,000 people die in U. S. from food-
borne illness. Some die from Trichinella in pork.
Millions of others suffer short term illness.
Irradiated pork is a safe and reliable way to eliminate
this pathogen. Process has been used successfully in
20 countries since 1950.
The negative description

Pork is exposed to radioactive materials, receives
300,000 rads of radiation--the equivalent of 30
million chest x-rays. Results in radiolytic products
in food. Some radiolytic products are carcinogens,
linked to birth defects. Process developed in 1950s
by Atomic Energy Commission.
Irradiated pork study:
Implications
1. Given positive descriptions of GM
foods, consumers initially will:
BUY THEM
2. Given negative descriptions of GM

foods, consumers initially will:
NOT BUY THEM
Irradiated pork study:
Implications
3. Given positive and negative descriptions of GM foods,
consumers initially will:
NOT BUY THEM
(if they have to pay more)

Lessons:
1. Both sides should be scrupulous in

providing reasons for all of their claims,
but especially for any negative claims.
Especially negative claims about our
opponents.
An ethical dilemma?
1. GM food appears to be safe.
2. GM food appears to be of great potential
benefit to millions of poor children.
3. Widespread anti-GM sentiment could
deprive children of GM foods.
Conclusion
If we are to overcome the all-or-nothing rhetorical trap we
must:
1. Be more careful with our negative

claims;
2. Be more careful with our rhetoric;
3. Be more creative in finding a third
way.
Intrinsic objections to ag biotech
1. Dont play God
Counter-examples:
High tech medicine
God wants us to
genetically engineer
food
4. Dont tinker with

nature
Counter-examples:
GIFT and in vitro
Plant cuttings
3. Dont change the

world
Counter-example:
Agriculture
4. Dont cross
species
Mules
Wheat
4. Dont cross species
Triticale
A cross of durum WHEAT
(Triticum durum ) and
RYE
(Secale cereale)
Ordinary wheat
is a hexaploid: a cross
of three separate species
has pieces of chromosomes from several

other species added to it
has a genome that is artificially

maintained and a natural method of
dissemination that has been disabled
Natural species barriers
are not so natural
DNA sequences of viral origin in

plants
DNA sequences of bacterial origin in

plants
Radiation Breeding
New:
- spaghetti
- whisky
- ...
~1800 new plants

Proponents of GM foods
Intrinsic arguments
Are not convincing.
Extrinsic arguments
GMOs are ethically acceptable
because benefits outweigh risks.
Proponents of GM foods
1. GM foods appear to be safe
2. U. S. consumers and environment are

adequately protected from risks of
allergens, toxins, and transgenes
3. Increased efficiency of production

needed in developing countries
1. GM foods appear to be safe
Genetic engineering of microbes
Recombinant Chymosin
Enzyme used for cheese
Originally from calf stomach
Bovine gene expressed in
GRAS microbes
FDA approved 1990
Now used in 70% of U.S.
cheese
GM foods appear to be safe
Recombinant amino acids
Aspartame
Artificial sweetener
Made from aspartic acid
and phenylalanine
Used in 5,000 products
Monosodium glutamate
Recombinant alpha amylase
Used to make HFCS

GRAS status in 1995
10% U.S. corn crop
processed into syrups
GM foods appear to be safe

1. Growing population
2. Declining gains in plant

breeding productivity
Extrinsic objections
Unsafe for environment?
Herbicide resistance - canola gene

flows into weedy relatives
Bt toxin kills monarch butterfly
larvae
Ethical Objections to Genetically
Modified
Foods
August 22, 2000
New Study Links

Biotech Corn to
Butterfly Deaths
By Carol Kaesuk Yoon
September 26, 2000
Biotech Corn Isn't
Serious Threat to
Monarchs, Draft U.S.
Report Finds
By Carol Kaesuk Yoon
Intrinsic objection:
Do not change the world (CW)
Counter-intuitive result:
We should have remained

hunter-gatherers.
(Comstock slide)
Medicine v. Food
Positive attitudes toward
biomedical uses
Less positive attitudes

toward food production uses
Two final questions
1. For the pro-GM movement
Why biotech? Why not alternative

methds of producing more food?
Alternative, non-GM, ways to
feed the world
Plant two rice
varieties instead
of one and
Double rice
production by
Reducing losses
to a fungus, blast.
Two final questions
2. For the anti-GM movement
What gives you (in the developed

world) the right to take the choice of
using biotech away from me (in the
developing world?
Helping improve
peoples
understanding of the
products of
agricultural and food
biotechnology.
CBI
Agriculture itself is unnatural
Many things which are natural are
harmful to man
Snake, spider toxins
Some bacteria, viruses
Plant and mycotoxins
In tearing up the land for agriculture, were
doing something unnatural, as Comstock
points out:
What is Bioinformatics?
Bioinformatics = Biology + Computer Science
Main Goal: To discover new biological insights
Post-Genomic Era:
Comparative Genomics,
Functional Genomics,
Structural Genomics,
Proteomics,
Medical Informatics,
Pharmacogenomics,
Pharmacogenetics
Why is Bioinformatics
Important?
New laboratory technology allows us to collect
data faster than we can interpret it
Leads to a better understanding of basic biological
functions and processes
Capitalize on existing and emerging technologies
such as database-mining and distributed
computing
http://folding.stanford.edu/science.html
Bioinformatics Skills
Molecular Biology
Central Dogma of Molecular Biology
Experience with at least one or two
molecular biology software packages
Basic understanding of how to use
Unix/Linux shells
C/C++ or Java, and, Perl or Python
programming experience
NCBI On-Line Resources
Entrez
BLAST
Taxonomy Browser
Macromolecular 3D Structure Database
Other Resources
TIGR
Genome Projects
http://www.tigr.org/tdb/
European Bioinformatics Institute
ClustalW
http://www.ebi.ac.uk/clustalw/
SWIS-PROT
http://www.ebi.ac.uk/swissprot/index.html
Swiss Institute of Bioinformatics
ExPASy
http://ca.expasy.org/
Protein Data Bank
http://www.rcsb.org/pdb/
NCBI
National Center for Biotechnology Information
http://www.ncbi.nih.gov
Conducts research on fundamental biomedical problems at the

molecular level using mathematical and computational methods
Fosters scientific communication by sponsoring meetings, workshops,
and lecture series
Develops, distributes, supports, and coordinates access to a variety of
databases and software for the scientific and medical communities,
and
Develops and promotes standards for databases, data deposition and
exchange, and biological nomenclature
NCBI Home page
Bioinformatics Example
Drosophila melanogaster (fruit fly)
model system for studying the development of animals from
embryo to adult
have gene called eyeless
Humans have a gene responsible for aniridia (mutated
PAX6 gene)
Causes the eyes to develop without irises
Insertion of PAX6 into eyeless D. melanogaster, normal
eyes develop
Sequence comparison can be used to gain insight into how eyeless
and aniridia work together
Entrez
NCBI Tutorials
http://
www.ncbi.nlm.nih.gov/Education/index.htm
l
Our Corporate
Members
Our Corporate
Members
Biotech Job opportunities
High Paying Jobs In India
Jobs in Delhi/NCR, Mumbai, Chennai,
Bangalore, Hyderabad & More
www. yellojobs. com/Jobs_ in_India
Nano Technology Jobs Site
Job Posting Site dedicated to
Nanotech Science and Engineeing
www. nano- career.com
Biotechnology
New Chemical Engineering Journal
Free online access to end 2008
www. apjChemEng.com
Employment Opportunities
MNC Companies Are Hiring
Post Resume & Get Placed Free!
ClickJobs. com/Employment/Post-CV
Internet Jobs in India
I Earn Rs.2000 Per Day in Part Time
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Single Step Resume Uploading
More Vacancies. Faster Response
www.JobsForYou.com
ScienceJobs.Org
Find Science Jobs and career info
at ScienceJobs.Org
www.ScienceJobs.Org
Job in india
Apply to Companies Directly.
Submit Your Resume Now!
JobStreet.com
Success Story of BIOCON
Biocon is India's leading biotechnology enterprise
Over the past 25 years, they have evolved from an small enzyme manufacturing
company to a fully integrated biopharmaceutical enterprise, focused on healthcare
At Biocon, success has been their ability to develop innovative technologies and
products and to rapidly leverage them to adjacent domains
This unique "integrated innovation" approach has yielded a host of patented

products and technologies that have enabled multi-level relationships with their
global clientele
As India's first and leading biotechnology company, Biocon extends its support to
numerous community outreach and corporate citizenship initiatives with special
concentration in the areas of healthcare, education and environment
The Biocon Foundation, set up in 2004 has recently launched Arogya Raksha
Yojana, a unique health initiative for rural India
279
Success Story of RELIANCE
The Reliance Group founded in 1932, is India's largest business house with
total revenues of over US $ 22.6 b and exports of US $ 3.6 b
It began as a an SME
The Group's activities span exploration and production of oil and gas,
refining and marketing, petrochemicals, textiles, financial services, insurance,
power, telecom and infocom initiatives
The Group exports its products to more than 100 countries the world over
Reliance Group revenue is equivalent to about 3.5 % of India's GDP
The Group contributes nearly 10 % of the country's indirect tax revenues and
over 6% of India's exports
Today, the Reliance Group has one of the largest family of shareholders in the
world
280
10,000 year old ancient civilization
325 languages spoken 1,652 dialects
18 official languages
29 states, 5 union territories
3.28 million sq. kilometers - Area
7,516 kilometers - Coastline
1.3 Billion population.
5600 dailies, 15000 weeklies and 20000
periodicals in 21 languages with a combined
circulation of 142 million.
GDP $576 Billion. (GDP rate 8%)
Parliamentary form of Government

Worlds largest democracy.
Worlds 4th largest economy.
World-class recognition in IT, bio-

technology and space.
Largest English speaking nation in the
world.
3rd largest standing army force, over
1.5Million strong.
nd
India: Technology Superpower
Geneva-based STMicroelectronics 15 of the world's major Automobile
is one of the largest semiconductor makers are obtaining components
companies to develop integrated from Indian companies.
circuits and software in India.
This business fetched India $1.5
Texas Instruments was the first Billion in 2003, and will reach $15
to open operations in Bangalore, Billion by 2007.
followed by Motorola, Intel,
Cadence Design Systems and
several others. New emerging industries areas
include, Bio-Informatics, Bio-
80 of the Worlds 117 SEI CMM Technology, Genomics, Clinical
Level-5 companies are based in Research and Trials.
India.
World-renowned TQM expert
5 Indian companies recently Yasutoshi Washio predicts that Indian
received the globally acclaimed manufacturing quality will overtake
Deming prize. This prize is given that of Japan in 2013.
to an organization for rigorous
total quality management (TQM)
practices. McKinsey believes India's revenues
from the IT
industry will reach $87 Billion by
2008.
Flextronics, the $14 billion

India: Pharmaceuticals
The Indian pharmaceutical industry at $6.5 billion and
growing at 8-10% annually, is the 4th largest
pharmaceutical industry in the world, and is expected to
be worth $12 billion by 2008.
Its exports are over $2 billion. India is among the top five bulk
drug makers and at home, the local industry has edged out
the Multi-National companies whose share of 75% in the
market is down to 35%.
Trade of medicinal plants has crossed $900M already.
There are 170 biotechnology companies in India, involved

in the development and manufacture of genomic drugs, whose
business is growing exponentially.
Sequencing genes and delivering genomic information

India: Technology Superpower
Over 100 MNCs have set up R&D facilities in India in the
past five years. These include GE, Bell Labs, Du Pont,
Daimler Chrysler, Eli Lilly, Intel, Monsanto, Texas
Instruments, Caterpillar, Cummins, GM, Microsoft and IBM.
Indias telecom infrastructure between Chennai, Mumbai

and Singapore, provides the largest bandwidth
capacity in the world, with well over 8.5 Terabits
(8.5Tbs) per second.
With more than 250 universities, 1,500 research

institutions and 10,428 higher-education institutes, India
produces 200,000 engineering graduates and another
300,000 technically trained graduates every year. (note: per
capita numbers are lower in comparison with first world, Russia and Israel,
indicating India should increase the number of educational institutions and
educational opportunities to its 1.3 billion population)
Besides, another 2 million graduate in other areas in

Indians abroad
A snapshot of Indians at the helm of leading Global
businesses
The Co-founder of Sun Microsystems (Vinod Khosla),

Creator of Pentium Chip (Vinod Dahm),
Founder and creator of Hotmail (Sabeer Bhatia),
Chief Executive of McKinsey & Co. (Rajat Gupta)
President and CFO of Pepsi Cola (Indra Nooyi)
President of United Airlines (Rono Dutta)
GM of Hewlett Packard (Rajiv Gupta)
President and CEO of US Airways (Rakesh Gangwal)
Chief Executive of CitiBank (Victor Menezes),
Chief Executives of Standard Chartered Bank (Rana Talwar)
Chief Executive officer of Vodafone (Arun Sarin)
President of AT & T-Bell Labs (Arun Netravali)
Vice-Chairman and founder of Juniper Networks (Pradeep Sindhu)
Founder of Bose Audio (Amar Bose)
Founder, chip designer Cirrus Logic (Suhas Patil )
Chairman and CEO of Computer Associates (Sanjay Kumar)
Head of (HPC WorldWide) of Unilever Plc. (Keki Dadiseth)
Chief Executive Officer of HSBC (Aman Mehta)
Director and member of Executive Board of Goldman Sachs (Girish Reddy)
Indians in the USA.
Statistics that show:
38% of doctors in the USA,

12% of scientists in the
USA,
36% of NASA scientists,
34% of Microsoft
employees,
28% of IBM employees,
17% of INTEL scientists,
13% of XEROX employees, US H1-B Visa
applicants
are Indians. country
of origin
1. India 44%
2. China 9%
Of the 1.5M Indians living in the USA, 1/5 of them live in
th
3. Britain 5%
the Silicon Valley. 4. Philippines
3%
5. Canada 3%
35% of Silicon Valley start-ups are by Indians. 6. Taiwan 2%
7. Japan 2%
8. Germany
Indian students are the largest in number among foreign 2%
Mahatma Gandhi
(1869-1948):
Gandhi was once asked what he thought about Western

Civilization. His response was: "I think it would be a
good idea.
"The greatness of a nation and its moral progress can
be judged by the way its animals are treated.
You must not lose faith in humanity. Humanity is an
ocean; if a few drops of the ocean are dirty, the ocean
does not become dirty.
The only devils in this world are those running around
inside our own hearts, and that is where all our battles
should be fought.
If all Christians acted like Christ, the whole world

would be Christian.
Woman, I hold, is the personification of self-sacrifice,
but unfortunately today she does not realize what
Sir C.V. Raman, (1888 1970)
1930 - Nobel Laureate in Physics for work on scattering of
light and Raman effect.
Sir Jagdish Chandra Bose, (1858 1937)

USA based IEEE has proved what has been a century old
suspicion amongst academics that the pioneer of wireless-
radio communication was Professor Jagdish Chandra
Bose and not Guglielmo Marconi.
Satyendranath Bose, (1894-1974)

Indian Physicist, who solved one of the mysteries of
quantum mechanics, showing that in the quantum world
some particles are indistinguishable. His collaborations
with Albert Einstein led to a new branch on statistical
mechanics know commonly known as the Einstein-Bose
statistics.
Srinivasa Ramanujam,(1887 1920):
Great Indian Mathematician, whose interest from
academics at Trinity, College, Cambridge, led him to
collaborate there and postulate and prove well over
3,542 theorems.
Subramanyan Chandrasekhar,(1910-
1995):
1983 Nobel Laureate in Physics. His many contributions
to physics, on the structure and evolution of stars including
rotational figures of equilibrium, stellar interiors, black holes,
radiative transfer, hydromagnetic stability, stellar dynamics.
Har Gobind Khorana, (b-1922 ):

1968 - Nobel Laureate in Medicine for work on
interpretation of the genetic code . Currently residing
as professor at MIT.
Amartya Sen, (b-1933):
1998 - The Nobel Prize for Economics for his redefining
work on ethical welfare economics. Currently residing as
Lamont University Professor Emeritus at Harvard, after
stepping down from the prestigious post of Master of Trinity
Ex-Prime Minister,
Sri Atal Bihari Vajpayee
A treaty was signed on 6 January, 2004, establishing a
South Asian Free Trade Area among the seven
SAARC countries (India, Pakistan, Bangladesh,
Bhutan, Nepal, Sri Lanka and Maldives) in the
Indian Continent.
India committed to a South Asian Union as the

ultimate objective, with mutual security
cooperation, open borders and a single currency in
Southern Asia in the long run.
"The bonds of ethnicity and culture which hold

together the peoples of this region are more
enduring than the barriers of political
prejudice that have been erected quite
recently.
.Friends, India is ready to do everything that is
Dr Abdul Kalam, President of India,
father of Indias space, missile and satellite programme and author of
India 2020 Vision.
I have three visions for India.
1.
In 3000 years of our history people from all over the world have
come and invaded us, captured our lands, conquered our minds. From
Alexander onwards. The Greeks, the Turks, the Moguls, the
Portuguese, the British, the French, the Dutch, all of them came and
looted us, took over what was ours.
Yet we have not done this to any other nation. We have not conquered
anyone. We have not grabbed their land, their culture, their history
and tried to enforce our way of life on them.
Why?
Because we respect the freedom of others. That is why my first vision
is that of FREEDOM.
I believe that India got its first vision of this in 1857, when we started
the war of independence. It is this freedom that we must protect and
nurture and build on. If we are not free, no one will respect us.
2.
My second vision for India is DEVELOPMENT. For fifty years we have
been a developing nation. It is time we see ourselves as a developed
nation. We are among top 5 nations of the world in terms of GDP. We
have 10% growth rate in most areas. Our poverty levels are falling.
Our achievements are being globally recognized today. Yet we lack the
self-confidence to see ourselves as
a developed nation, self-reliant and self-assured.
3.
Indias population to be the largest in the world
India is set to overtake China as the world's most

populous nation by 2050.
Indias population is expected to grow from 1.08bn to

1.63bn people, overtaking China, which is forecast to
reach 1.44bn from 1.3bn currently.
India, will also have the highest working population in the

World 700 million people out of 1.1 billion people are
young; the young population will continue till 2050.
Bio-forecast
$1500m
Indian Biotech Products & Services:
R&D Products & Services

Ag Biotech
Industrial Biotech
$750m
Medical Biotech
$ 150
$ 150
$150m $ 100
$ 350
2002
2005
The Indian Biotech business stands at US$150 m today... 2007
and is expected to be $ 750 m by 2005 and $1500 m by 2007
#3
Bio-clusters
DBT
PUNE UNIVERSITY NII
ALFA LAVAL CBT
PRAJ INDUSTRIES DABUR
SERUM INSTITUTE GENOMED
WOCKHARDT PANACEA BIOTECH
MIDC BIOTECH JUBILANT
PARK
New Delhi
IISC
CCMB
NCBS
JNCASR BHARAT BIOTECH
IBAB Pune SHANTHA BIOTECH
ASTRA ZENENCA
AURIGENE DRL
AVESTHAGEN Hyderabad BIOLOGICAL E
BANGALORE GENEI
BIOCON, SYNGENE KNOWLEDGE PARK
GANGAGEN S-P BIOTECH PARK
JUBILANT BIOSYS
METAHELIX Chennai TCS
SARTORIUS
STRAND GENOMICS
BIOTECH PARK Bangalore
CROs: CLINIGENE, CLINITEC,
LOTUS LABS ANNA UNIVERSITY
MADURAI KAMARAJ
UNIVERSITY
SPIC #5
Bio-resources
Human resources ~ large English speaking skill base: 3 million graduates,

700,000 post graduates & 1500 PhDs have qualified in bio-sciences and
engineering.
Biotech Industry: Manpower
#8
Bio-resources
Low cost of innovation ~ employment costs of post graduates and PhD level
scientists and engineers is a fraction of those in Europe and USA.
Lucrative policies & incentives ~ from center and state governments. 150 per
cent weighted tax deduction applicable for companies investing in research
and development (R&D) activity.
Diverse skill sets ~ Ranging from biologists, geneticists, medical
practitioners,clinicians, paramedics, statisticians, informaticians,
biophysicists, chemists, pharmacologists, computer scientists and software
engineers.
Supportive Regulatory regime ~ Approval of Bt cotton for commercial
cultivation paves way for other GM crops.
3 recombinant biologicals approved. Hep B vaccine, EPO and Interferon Alpha
2b.Stem cell research is an approved domain.
#9
Bio-Processes in India
Indias well developed fermentation and purification skills are being

leveraged to a globally competitive bioprocessing platform...
VACCINES RECOMBINANT MONOCLONAL

THERAPEUTICS ANTIBODIES
Serum Institute Biocon Biocon

Shantha Biotech Wockhardt Shantha Biotech
Bharat Biotech Dr. Reddys
Wockhardt Shantha Biotech
Panacea Biotech Bharat Biotech
Dr. Reddys
Hep B, Human Insulin,

Combi-vaccines EPO, GCSF, SK, HGH, Enti-EGF & others
Rabies, Leprosy Interferon 2B
#15
CLINICAL DEVELOPMENT
India is fast emerging as the hotbed of clinical research. One new CRO is being
created every month...
NEW
BIO BIOMETRICS/ DATA SITE
DRUG
STUDIES MANAGEMENT MANAGEMENT
TESTING
Lambda Quintiles Neemans

Siro Clinpharm
Clinigene Clinigene GSK
Quintiles Lotus Labs Pfizer
Synchron Vimta Labs Novartis
Pfizer Apothecaries Eli Lilly
Clintec Synchron DiagnoSearch
Covance Wellquest Clinworld
Eli Lilly Infotech
Novartis
AstraZeneca
#19
R&D ORGAINZATIONS
ORGANIZATION NUMBER STRENGTH
1. CSIR
LABS AND RES. 118 SCIENTISTS 10, 934
2. ICAR
AGRI. UNIV. 43 SCIENTISTS 6,281
R&D INST. 89
3. ICMR
R&D INST./CENT. 26 SCIENTISTS 732
Cont.//
4. DBT
R&D INST. 8 SCIENTISTS 700
5. DRDO
R&D INST. 52 SCIENTISTS 6,500
6 DAE.
R&D INST. 17 SCIENTISTS 5,000
7. DOS
ISRO SCIENTISTS 10,000
INSTITUTIONS OF HIGER EDUCATION (2001)
1. HIGHER EDUCATION NUMBER STRENGTH
UNIVERSITIES 253 FACULTY 411,628
COLLEGES 13,150 STUDENTS 8,399,433
SC STUDENTS 1,670,273
2. ENGINEERING EDUCATION
HIGHER EDUCATION &RES.
IITs 7 FACULTY 2,500
STUDENTS 23,000
ENGG.COLLEGES STUDENTS 463,965
DEGREE 1347 DEGREE 262,882
DIPLOMA 1228 DIPLOMA 201,093
Cont.//
3. MEDICAL EDUCATION
MEDICAL COLLEGES 189 STUDENTS 18,168
PG STUDENTS 3,181
PG DIPLOMA 1,316
BIOTECHNOLOGY IN INDIA
SKILLED MANPOWER
GRADUATES 3,000,000
POST-GRADUATES 70,000
Ph.Ds 1,500
CONVENTIONAL BIOTECH. COMPANIES 110
MODERN BIOTECH. COMPANIES 60
BIOTECH ASSOCIATED INSTITUTIONS
RESEARCH INSTITUTEIONS 241
INDUSTRIES 328
Cont.//
CONSUMPTION OF BIOTECH.
PRODUCTS IN 1999 US $ 1.79 BN.
PROJECTED CONSUMPTION IN 2005 US $ 3.72 BN.

PROJECTED CONSUMPTION IN 2010 US $ 4.27 BN.
MAIN ACTITIVITES AND POTENTIAL
RICH BIODIVERSITY-INDIA HAS 7.76% OF TOTAL

MAMMAL SPECIES 12.6% OF BIRD SPECIES,
11.7% OF FISH AND ROUGHLY 6.0% OF TOTAL
FLOWERING PLANTS PRESENT IN THE WORLD
WITH EXCELLENT AGRO-CLIMATIC
CONDITIONS. THIS IS A GREAT RESOURCE FOR
RESEARCH AND DEVLOPMENTAL ACTIVITIES IN
BIOTECHNOLOGY.
FOCUS GENOMICS, RECOMBINANT DNA,
TRANSGENICS. STEM CELL RESEARCH AND
NEW DRUGS DEVELOPMENT.
Cont.//
VACCINE MARKET AROUND US $ 100 Mn.
VALUED
GROWTH 20%
RATE
DIAGNOSTICS MARKET DEMAND US $ 50 Mn.

DEMAND GROWING RAPIDLY
ANIMAL HEALTH BIOTECHNOLOGY MARKET
RAPIDLY GROWING
ENZYMES 70% REQUIREMENT IMPORTED
MARKET GROWING WITH
INCREASING AWARENESS
Cont.//
PHARMA TODAY 40-45% OF ALL DRUG ORIGINATE
INDUSTRY IN BIO-TECHNOLOGY BUT IN THE NEXT
DECADES BIOTECHNOLOGY WILL HAVE
A MUCH GREATER ROLE ON THE PROD-
UCTION OF DRUGS AND PHARMACUET-
CALS.
FERMENTATION MODEST FERMENTATAION INDUSTRY
INDUSTRY MAINLY BREWERIE,
ANTIBIOTICS AND CHEMICALS
BUT GREAT POTENTIAL IN THE
AREAS OF FOOD PROCESING,
PHARMA AND CHEMICALS.
Cont.//
FOOD INDIA IS THE LARGEST PRODUCER
PROCESSING OF MILK AND SECOND LARGEST
INDUSTRY PROCUCER OF FRUITS, VEGETABLES
AND FOODGRAINS. BUT PROCESSING
IS LIMITED TO ONLY 2% OF FRUITS AND
VEGETABLES AND 15% MILK. THUS,
TREMENDOUS SCOPE FOR THE
DEVELOPMENT OF PROCESSING OF MILK
FRUITS &VEGETABLES, MEAT, FISH AND
FOODGRAINS
Cont.//
AGRI-BIOTECH. -GENETICALLY ENGINEERED
SEEDS AND PLANTS HAVE A
MARKET OF US $ 250 Mn..
-DEVELOPMENT OF TRANSGENICS
FOR QUALITY AND CHARACTERS.
-DEVELOPMENT OF TRANSGENICS
WITH RESISTANCE TO DROUGHT
AND OTHER ABIOTIC STRESSES.
-TISSUE CULTURE FOR
PROPAGATION OF PLANTS.
-DEVELOPMENT OF MEDICINAL AND
OTHER AROMATIC PLANTS.
ADVANTAGE INDIA
NO INDUSTRIAL LICENCE REQUIRED FOR FOODS
EXCEPT ALCHOHLIC PRODUCTS.
NATIONAL POLICY TO INCREASE FOOD PROCESSING
FROM PRESENT 2% TO 10% BY 2010.
INVESTMENT OPPORTUNITIES:
UPTO 51% FOREIGN EQUITYAUTOMATIC APPROVAL.
UPTO 100% EQUITY ALLOWED ON APPROVAL UNDER
FOREIGN DIRECT INVESTMENT (FDI) SCHEME EXCEPT
MALTED FOOD, ALCHOHOLIC BEVERAGES.
CUSTOM DUTIES SUBSTANTIALLY REDUCED ON PLANT
AND EQUIPMENT, RAW MATERIALS AND
INTERMEDIARIES.
INDIA IN THE NEW KNOWLEDGE ECONOMY
Healthcare
Pharma & Biotechnology
Information & Communication

Technology (ICT)
Drugs & Pharmaceuticals
Indias strength in information technology, biology and

synthetic chemistry presents opportunities for research
services in areas like bio-informatics, structure based drug
design and synthetic and medical chemistry.
Development services like clinical trials and data
management services have similar potential.
With these new strengths Indian pharmaceutical industry
is expected to grow from $ 6 billion in 2001 to $21 to $ 25
billion by 2010.
Biotechnology
Large expertise exists for research and development in

the field of genomics, bio informatics, DNA technologies,
clinical studies and genetically modified crops.
Stem cell biology, molecular probes, genetic based
vaccines and molecular taxonomy are taking off.
Indian investment in bio technology is expected to
increase five fold from $ 2 billion to $ 10 billion during the
decade.
1. THE HUMAN CAPITAL EDGE
Over 3 million scientific & technical manpower

Stock of over 0.8 million post graduates in science.
Over 1 million graduate engineers
0.4 million doctors
0.3 million graduates in agriculture and veterinary
sciences.
Today India turns out more than 50,000 computer
professionals and 360,000 engineering graduates
each year.
2. SIZE OF DOMESTIC MARKET
1.1 billion population
Consuming middle class

population
300 million
Size of the market at Purchasing

Power Parity (PPP) US$ 1.5
trillion
RESEARCH & DEVELOPMENT
More than 100 global companies outsource R&D
facilities from India
GE John F Welch Technology Centre Companys largest
research outfit outside the US
GE Medical Systems India as sole sourcing base for its
portable ultrasound scanner
Monsanto First non-US research facility
Eli Lilly largest research facility in Asia and 3rd largest in the
world
Texas Instruments Digital Signal Processor developed in
India controls 50% of the world market
AVL, Austria India as base to do R&D for the companys
Japan centre
Emerging skill shortages in the Indian industry
Analysis of some major sectors
BIOTECHNOLOGY
SKILL GAP AREAS :
Scientists & Researchers : 80% Shortage
Production staff: 18% Shortage
Legal and Regulatory experts: 22%
Industry Comments
Companies have reported that on an average they have to spend Rs 2- 6 lacs per person per
year on the training of candidates possessing Masters Degree in Biotechnology, Organic
Chemistry / Biology.
According to industry representatives the government should put a lot of emphasis on
increasing qualified professionals (particularly doctorates) in the following areas molecular
biology, biochemistry, biotechnology, assay development biology, medicinal chemistry,
analytical chemistry, synthetic organic chemistry.
INTERACTIONS WITH GREAT
NATIONAL LEADERS
I cannot forget the services rendered by
commercial class, but I want you to make
Congress your own and we would willingly
surrender the reins to you. The work can be
better done by you. But if you decide to assume
the reins, you can do so only on one condition.
You should regard yourselves as trustees and
servants of the poor. Your commerce must be
regulated for the benefit of the toiling millions
MAHATMA GANDHI
Speaking at Annual Session on 7th
April,1931
Achievements
~ 50 Biotech PCT Patents have been filed in India
~ 20 r-DNA product clinical trials are on-going in India
~ 10 r-DNA products have been approved in India
There are 6 r-DNA products being manufactured in India

(Hep B, Interferon, GCSF, EPO, SK, BT cotton)
There are 6 companies manufacturing recombinant vaccines
There are 2 companies that will manufacture r-Human Insulin
There are 2 companies developing Monoclonal Antibodies
There are ~25 companies marketing Biotech products
#4
The India Advantage
Excellent network
Well-developed
of research laboratories
base industries
Proficiency in Pro active

English policy framework
Extensive clinical trial

opportunities
Rich biodiversity
Low manpower Trained manpower

costs and knowledge base
320
Opportunity in various Industry Sectors
Software
Biotechnology
Outsourcing
Retailing
Real Estate
Auto Components
Telecommunication
Media & Entertainment
321
Opportunities in Biotechnology
Biotechnology is a fast emerging sector in India
The consumption of biotech products in India was US $ 1.8 b

in 1999 and is expected to cross US $ 4.2 b by 2010
It is estimated that 10 % of the researchers and 15% of the

scientists in pharmaceuticals and biotech R&D in USA are of
Indian origin
India is churning out 2 m graduates, 300,000 post

graduates and 1,500 PhD's, every year
Biocon, the first biotech company was launched in 1978
Indian Firms such as Shantha Biotech, Bharat Biotech,

Dr. Reddy's Lab and Biocon are all very successful firms
322
Biotech Market - A consistent Uptrend
25 % growth in investment
70 % growth in employment
2002- 03
74 % growth in R&D
manpower
Size 2003 - 04 US $ 705 m
US $ 5 b annual
revenues
2010
1 m skilled jobs
10 % of global industry
Source: Confederation of Indian Industry (CII) 323
Biotechnology job India - Developing a career in
biotechnology in ...
Jobs in biotechnology field from Biotech Desk India
that offers exciting careers
in biotech field ... Career in Biotechnology -
Biotechnology jobs in India ...
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Biotechnology, biotech jobs, careers in
biotechnology ...
biotechnology software, biotechnology consulting jobs.
India, the darling of the
world as far as bio technology sector is concerned offers
tremendous ...
www.123biotech.com/
BIOINFORMATICS INSTITUTE OF INDIA
:-:-:- Bioinformatics Jobs & Careers
Bio Career Center, Search jobs and post your
rsum, courtesy of the Biotechnology
Industry Organization, a worldwide trade group.
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www.bii.in/page/careers/job_career.asp
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Biotechnology is an interdisciplinary
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www.careerage.com/resources/career_choices/bi
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Pharmaceutical Careers
Pharmaceutical Science & Technology Premier Pharmaceutical Institute
www.tipt.com
Life Science Find Great Life Science Resources Here.
Learning.TitanFacts.com
Biotechnology Find vendors and businesses in our business directory.
www.allbusiness.com
ScienceJobs.Org Find Science Jobs and career info at ScienceJobs.Org
www.ScienceJobs.Org
Phenomenome Profiler
The first universal mass spec software designed for metabolomics.
www.phenomenomeinformatics.com
Crystalization
Looking for Crystalization in pharmaceutical applications?
www.fuji-research.com
PubMed Medline
Search 17 million+ articles FREE Access toConference Abstracts
www.biowizard.com
Laboratory Consultants
University, Industry & Government Research Facilities Design
www.rfd.com
Global LifeScience Events
Senior level conferences for the worldwide Life Science industry
www.terrapinn.com
Mass Spectrometry
Ion Traps, TOF & FTMS Bruker Daltonics Life Science Tools
www.bdal.de
Biocon is a research-driven, global healthcare company
with a strong matrix of capabilities along the
biopharmaceutical value chain. Focusing on unmet
medical needs in cancer, diabetes and inflammatory
diseases, we offer novel therapies on a platform of
affordable innovation.
We believe our strategic licensing partnerships will rapidly
provide us wider global access and greater market
penetration. More significantly, they will enable us to
deliver breakthrough therapeutics to millions of patients
the world over.
INNOVATION WITHOUT AFFORDABILITY AND
ACCESSIBILITY IS LIKE
THERAPY WITHOUT PATIENTS.
2007, Biocon. All Rights Reserved

Some Indian Examples Benchmarked with the
Best In The World
329
Where is happiness !!!!!!
WHERE IS HAPPINESS !!!!!!!!
Bags of Gold can never buy
The happiness your hunting for--(2)
happiness is the money you have
nor the gold nor the world
nor the cigar you smoke
he is loving Jesus everyday
Thanks for your attention
ANY QUESTIONS
THANK YOU
THANK YOU
Please contact me at:

email: annaduraipillai@yahoo.co.in
telephone: 0416-2252255,
9443035147
Rethinking Scholarly Publishing New
Models of Publishing
New technologies solving old problems BioMed Central
of scholarly publishing http://www.biomedcentral.com
an independent online publishing house
provides free access to peer-reviewed biological and medical research it publishes
promotes rapid and efficient communication of science
Old Model latest new journals include: Annals of Clinical Microbiology and Antimicrobials, Comparative Hepatology,
International Journal for Equity in Health, Journal of Negative Results in Biomedicine, Journal of Immune
Based Therapies and Vaccines
Author Library
Scholar
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$ $ Reader
Author/Scholar/Researcher Faculty of 1000
http://www.facultyof1000.com
$ $ $ $ over 1400 of the worlds leading scientists select and evaluate the best papers in biology
highlights papers on the basis of their scientific merit
unique organization and personalization features, including personalized email alerts
Journal Editor
Editorial Board
Peer Review
$ $ Commercial Publisher
offers an immediate rating of individual papers by the authors' peers
participating University of Utah authors: Brenda Bass, Kendal Broadie, Bradley Cairns, Mario Capecchi,
Gerald Gleich, Kent Golic, Janet Lindsley, Susan Mango, Alejandro Sanchez-Alvarado, Gerald Spangrude
SPARC
New is an alliance of 200 member institutions world-wide
supports development of vibrant and economically viable new models of scholarly communication
Model works with publishers whose business goals are compatible with SPARC
Author Library encourages competition so that journal subscriptions are reasonably priced and libraries can afford them
Scholar Reader creates systems that expand information dissemination and use in a networked digital environment
Researcher Author/Scholar/Researcher
facilitates improved and expanded communication by researchers
stimulates creation of better, faster, and more economically sustainable systems for distributing new
knowledge
PubMed Central
Journal Editor digital archive of life sciences journal literature
Editorial Board
not a journal publisher
Peer Review
access to PubMed Central (PMC) is free and unrestricted
managed by the National Center for Biotechnology Information at the National Library of Medicine
Jeanne Le Ber, Librarian

Spencer S. Eccles
Health Sciences Library
What is Biotechnology?
How about some definitions
General Definition
The application of technology to improve
a biological organism
Detailed Definition
The application of the technology to modify the
biological function of an organism by adding genes
from another organism
These definitions imply biotechnology
is needed because:
Nature has a rich source of variation
Here we see bean has many

seedcoat colors and patterns
in nature
But we know nature does not have

all of the traits we need
But nature does not contain all the
genetic variation man desires
Fruits with vaccines
Grains with improved nutrition

What controls this natural variation?
Allelic differences at genes control a specific trait
Definitions are needed for this statement:
Gene - a piece of DNA that controls the

expression of a trait
Allele - the alternate forms of a gene

What is the difference between
genes and alleles for Mendels Traits?
Mendels Genes
Plant height Seed shape
Smooth Wrinkled
Allele
Tall Short
Allele
This Implies a
Genetic Continuum
A direct relationship exists between the gene, its alleles,

and the phenotypes (different forms ) of the trait
Alleles must be:

similar enough to control the same trait
but different enough to create different phenotypes
Allelic Differences for Mendels Genes
Plant Height Gene
Gene: gibberellin 3--hydroxylase

Function: adds hydoxyl group to GA20 to make GA1
Role of GA1: regulates cell division and elongation
Mutation in short allele: a single nucleotide converts
an alanine to threonine in final protein
Effect of mutation: mutant protein is 1/20 as active
Allelic Differences for Mendels
Seed Shape Gene
Gene: strach branching enzyme (SBE) isoform 1

Function: adds branch chains to starch
Mutation in short allele: transposon insertion
Effect of mutation: no SBE activity; less starch, more
sucrose, more water; during maturation seed looses
more water and wrinkles
Central Dogma of Molecular Genetics
(The guiding principle that controls trait expression)
Protein Trait
(or phenotype)
Translation
Seed shape
DNA RNA
Transcription
(gene)
Plant height
In General, Plant Biotechnology Techniques
Fall Into Two Classes
Gene Manipulation
Identify a gene from another species which controls
a trait of interest
Or modify an existing gene (create a new allele)
Gene Introduction
Introduces that gene into an organism
Technique called transformation
Forms transgenic organisms
Gene Manipulation Starts
At the DNA Level
The nucleus
contains DNA
Source: Access Excellence

DNA Is Packaged
Double-stranded
DNA
is condensed
into
Chromosomes

Chromosomes Contain Genes
Chromosome
Gene

Genes Are Cloned Based On:
Similarity to known genes

Homology cloning (mouse clone used to obtain human gene)
Protein sequence
Complementary genetics (predicting gene sequence
from protein)
Chromosomal location
Map-based cloning (using genetic approach)
Homology Cloning
Clones transferred
to filter
Human clone
Mouse probe
library
added to filter
Hot-spots are human

homologs to mouse gene
Complementary Genetics
1. Protein sequence is related to gene sequence

NH3+-Met-Asp-Gly--------------Trp-Ser-Lys-COO-
ATG GAT-GCT TGG-AGT-AAA
C C C G
A TCT
G C
A
G
2. The genetic code information is used to design PCR primers

Forward primer: 5-ATGGAT/CGCN-3
Reverse primer: 5-T/CTTNC/GT/ACCA-3
Notes: T/C = a mixture of T and C at this position;

N = a mixture of all four nucleotides
Reverse primer is the reverse complement of the gene sequence
(cont.)
3. Use PCR to amplify gene fragment
a. template DNA is melted (94C)
3 5
5 3
3 5
5 3
b. primers anneal to complementary site in melted DNA (55C)

3 5
5 3
c. two copies of the template DNA made (72C)

3 5
5 3
PCR Animation
Denaturation: DNA melts

Annealing: Primers bind
Extension: DNA is replicated
PCR Again
(cont.)
4. Gene fragment used to screen library
Clones transferred
to filter
Human clone
library PCR fragment
probe added to filter
Hot-spots are human gene

of interest
Map-based Cloning
Gene Marker
1. Use genetic techniques to
find marker near gene
Gene/Marker
2. Find cosegregating marker
3. Discover overlapping clones

(or contig) that contains the marker Gene/Marker
Gene/Marker
4. Find ORFs on contig
5. Prove one ORF is the gene by Mutant + ORF = Wild type?

transformation or mutant analysis Yes? ORF = Gene
Gene Manipulation
It is now routine to isolate genes
But the target gene must be carefully chosen
Target gene is chosen based on desired phenotype
Function:
Glyphosate (RoundUp) resistance
EPSP synthase enzyme
Vitamin A biosynthetic pathway enzymes
The RoundUp Ready Story
Glyphosate is a broad-spectrum herbicide

Active ingredient in RoundUp herbicide
Kills all plants it come in contact with
Inhibits a key enzyme (EPSP synthase) in an amino acid pathway
Plants die because they lack the key amino acids
A resistant EPSP synthase gene allows crops

to survive spraying
RoundUp Sensitive Plants
Shikimic acid + Phosphoenol pyruvate
+ Glyphosate
X
Plant
EPSP synthase
Without amino acids,

X
3-Enolpyruvyl shikimic acid-5-phosphate
(EPSP)
plant dies
X
X
Aromatic
amino acids
RoundUp Resistant Plants
+ Glyphosate
RoundUp has no effect;
Bacterial enzyme is resistant to herbicide
EPSP synthase
3-enolpyruvyl shikimic acid-5-phosphate

(EPSP)
With amino acids,
plant lives
Aromatic
amino acids
The Golden Rice Story
Vitamin A deficiency is a major health problem
Causes blindness
Influences severity of diarrhea, measles
>100 million children suffer from the problem
For many countries, the infrastructure doesnt exist

to deliver vitamin pills
Improved vitamin A content in widely consumed crops

an attractive alternative
-Carotene Pathway in Plants
IPP
Geranylgeranyl diphosphate
Phytoene synthase
Phytoene
Problem: Phytoene desaturase
Rice lacks
these enzymes -carotene desaturase
Lycopene
Lycopene-beta-cyclase
Normal
Vitamin A -carotene
Deficient (vitamin A precursor)
Rice
The Golden Rice Solution
-Carotene Pathway Genes Added
IPP
Daffodil gene Phytoene synthase
Phytoene
Vitamin A
Phytoene desaturase
Pathway Single bacterial gene;
is complete performs both functions
and functional -carotene desaturase
Lycopene
Daffodil gene Lycopene-beta-cyclase
Golden -carotene
Rice (vitamin A precursor)
Metabolic Pathways are Complex
and Interrelated
Understanding pathways
is critical to developing
new products
Modifying Pathway Components
Can Produce New Products
Turn On Vitamin Genes =
Relieve Deficiency
Modified Lipids =
New Industrial Oils
Increase amino acids =

Improved Nutrition
Trait/Gene Examples
Trait Gene
RoundUp Ready Bacterial EPSP

Golden Rice Complete Pathway
Plant Virus Resistance Viral Coat Protein
Male Sterility Barnase
Plant Bacterial Resistance p35
Salt tolerance AtNHX1
Introducing the Gene or
Developing Transgenics
Steps
1. Create transformation cassette
2. Introduce and select for transformants

Transformation Cassettes
Contains
1. Gene of interest
The coding region and its controlling elements
2. Selectable marker
Distinguishes transformed/untransformed plants
3. Insertion sequences
Aids Agrobacterium insertion
Gene of Interest
Promoter TP Coding Region
Promoter Region
Controls when, where and how much the gene is expressed
ex.: CaMV35S (constitutive; on always)
Glutelin 1 (only in rice endosperm during seed development)
Transit Peptide
Targets protein to correct organelle
ex.: RbCS (RUBISCO small subunit; choloroplast target
Coding Region
Encodes protein product
ex.: EPSP
-carotene genes
Selectable Marker
Promoter Coding Region
Promoter Region
Normally constitutive
ex.: CaMV35s (Cauliflower Mosaic Virus 35S RNA promoter
Coding Region
Gene that breaks down a toxic compound;
non-transgenic plants die
ex.: nptII [kanamycin (bacterial antibiotic) resistance]
aphIV [hygromycin (bacterial antibiotic) resistance]
Bar [glufosinate (herbicide) resistance]
Effect of Selectable Marker
Non-transgenic = Lacks Kan or Bar Gene
Plant dies in presence

of selective compound
X
Transgenic = Has Kan or Bar Gene
Plant grows in presence

Insertion Sequences
TL TR
Required for proper gene insertions

Used for Agrobacterium-transformation
ex.: Right and Left borders of T-DNA
Lets Build A Complex Cassette
pB19hpc (Golden Rice Cassette)
TL aphIV 35S Gt1 psy 35S rbcS crtl TR
T-DNA Hygromycin Phytoene Phytoene T-DNA

Border Resistance Synthase Desaturase Border
Insertion Selectable Gene of Gene of Insertion

Sequence Marker Interest Interest Sequence
Delivering the Gene
to the Plant
Transformation cassettes are developed in the lab
They are then introduced into a plant
Two major delivery methods
Agrobacterium
Tissue culture
Gene Gun required to generate
transgenic plants
Plant Tissue Culture
A Requirement for Transgenic Development
Callus
grows
A plant part Shoots
Is cultured develop Shoots are rooted;
plant grows to maturity
Agrobacterium
A natural DNA delivery system
A plant pathogen found in nature

Infects many plant species
Delivers DNA that encodes for plant hormones
DNA incorporates into plant chromosome
Hormone genes expressed and galls form at infection site
Gall on
stem
Gall on
leaf
The Galls Can Be Huge
Natural Infection Process Is Complex
But Natures Agrobacterium
Has Problems
Infected tissues cannot be regenerated (via tissue culture)
into new plants
Why?
Phytohormone balance incorrect regeneration
Solution? Transferred DNA (T-DNA) modified by
Removing phytohormone genes
Retaining essential transfer sequences
Adding cloning site for gene of interest
The Gene Gun
DNA vector is coated onto gold or tungsten particles
Particles are accelerated at high speeds by the gun
Particles enter plant tissue
DNA enters the nucleus and

incorporates into chromosome
Integration process unknown

Transformation Steps
Prepare tissue for transformation

Tissue must be capable of developing into normal plants
Leaf, germinating seed, immature embryos
Introduce DNA
Agrobacterium or gene gun
Culture plant tissue

Develop shoots
Root the shoots
Field test the plants

Multiple sites, multiple years
The Lab Steps
Lab Testing The Transgenics
Insect Resistance Cold Tolerance
Transgene= Transgene=
Bt-toxin protein CBF transcription factors
More Modern Examples
Salt Tolerant Mercury Resistance
Glyoxylase I Mercuric ion reductase
The Next Test Is The Field
Herbicide Resistance
Non-transgenics
Transgenics
Final Test
Consumer Acceptance
RoundUp Ready Corn
Before After
The Public Controversy
Should we develop transgenics?
Should we release transgenics?
Are transgenics safe?
Are transgenics a threat to non-transgenic

production systems?
Are transgenics a threat to natural

eco-systems?
North Carolina
Biotechnology
Center
CFG Corporate Partner Examples
Science &
Technology
Technology
Development
Program
Science &
Intellectual Exchange Groups
Technology
Science &
THE MAKING OF A GMO CROP VARIETY
Backcrossing and selection (6- 8 generations)
Transgenic Commercial
line variety
Commercial Transgenic Line
Biotechnology
GMOs = Genetically Modified Organisms
Broadly defined: any microbe, plant, or animal developed through

breeding and selection
Narrowly defined: organisms produced by gene transfer techniques
Current examples of GMO Crops
herbicide-resistant crops insect-resistant crops

soybean cotton
corn potato
canola (rapeseed) corn
many others
GMO Crops on the Horizon

Corn, soy, canola with improved Nutraceuticals Golden Rice
Vaccines in plants
nutritional qualities for animal feed
Improved yields and stress tolerance
Crops with specialty starches and oils
for industrial processes
PLANT GENETIC ENGINEERING
Product Concepts and Technical Feasibility
Building the Transgenes
Plant Transformation
Event Selection
Plant Breeding
Seed Production and Marketing
Detection of GMO Crops in the Commodity Chain

ON/OFF Switch Makes Protein stop sign
PROMOTER INTRON CODING SEQUENCE poly A signal
Plant Transgene
Plant Selectable
Marker Gene
Plasmid DNA
Construct
bacterial genes
antibiotic marker
replication origin
Plant Transformation
The introduction and expression of
genes into plants is a three step process:
DNA Delivery to Target Cells
Selection and Regeneration
Event Selection
Plant Transformation DNA Delivery
microprojectile bombardment
Agrobacterium tumefaciens
biolistics or gene gun
natural property of
tiny DNA-coated particles are Agrobacterium to transfer DNA
shot into plant cells to host plant cells is exploited to
introduce genes of interest
versatile method
difficult with cereal crops
complex DNA integration
patterns: tandem arrays of simple DNA integration patterns
fragmented molecules
Plant Transformation Selection
At best only 1 in 1000 cells integrate tissue culture cells
under selection
delivered DNA
Transformed cells (events) are marked by
co-introducing gene that provides
resistance to selective agents
Transformed cells are selected by killing
non-transformed cells with selective agent.
Herbicide Leaf Paint Assay
Three main types of selective agents: transgenic non-transgenic
resistant susceptible
antibiotics
herbicides
plant growth regulators
Selectable markers assist in following

inheritance of transgenes.
GMOs: Why the Controversy?
Genetic engineering is a powerful new technology that is in general
poorly understood and whose long term effects are unknown.
GMOs are an innovation that have and will continue to impact all
facets of the global agricultural economy.
Production Processing
Commodity Handling Consumer Products

GMO Crops: Three Major Issues
1. Food safety and environmental impacts
2. Global trade
3. Increased corporate control of agriculture

Increased Corporate Control of Agriculture
The Development of GM Crops is Expensive

Intellectual property and patent protection
Consolidation/vertical integration increases ability to capture profits
Ag-biotech is a recent example of a century-old trend

Next Generation of Ag Biotech Products
Golden Rice increased Vitamin A content

(but not without controversy)
transgene = three pathway enzymes
Sunflower white mold resistance

transgene = oxalate oxidase from wheat
Source: Minnesota
Microscopy Society
DNA Is Packaged
Double-stranded
DNA
is condensed
into
Chromosomes

Chromosomes Contain Genes
Chromosome
Gene
Homology Cloning
Clones transferred
to filter
Human clone
Mouse probe
library
added to filter
Hot-spots are human

homologs to mouse gene
1. Protein sequence is related to gene sequence

NH3+-Met-Asp-Gly--------------Trp-Ser-Lys-COO-
ATG GAT-GCT TGG-AGT-AAA
C C C G
A TCT
G C
A
G
2. The genetic code information is used to design PCR primers

Forward primer: 5-ATGGAT/CGCN-3
Reverse primer: 5-T/CTTNC/GT/ACCA-3
Notes: T/C = a mixture of T and C at this position;

N = a mixture of all four nucleotides
Reverse primer is the reverse complement of the gene sequence
(cont.)
3. Use PCR to amplify gene fragment
a. template DNA is melted (94C)
3 5
5 3
3 5
5 3
b. primers anneal to complementary site in melted DNA (55C)

3 5
5 3
c. two copies of the template DNA made (72C)

3 5
5 3
Gene Manipulation
It is now routine to isolate genes
But the target gene must be carefully chosen
Target gene is chosen based on desired phenotype
Function:
Glyphosate (RoundUp) resistance
EPSP synthase enzyme
Vitamin A biosynthetic pathway enzymes
PCR Animation
Denaturation: DNA melts

Annealing: Primers bind
Extension: DNA is replicated
PCR Again
(cont.)
4. Gene fragment used to screen library
Clones transferred
to filter
Human clone
library PCR fragment
probe added to filter
Hot-spots are human gene

of interest
Map-based Cloning
Gene Marker
1. Use genetic techniques to
find marker near gene
Gene/Marker
2. Find cosegregating marker
3. Discover overlapping clones

(or contig) that contains the marker Gene/Marker
Gene/Marker
4. Find ORFs on contig
5. Prove one ORF is the gene by Mutant + ORF = Wild type?

transformation or mutant analysis Yes? ORF = Gene
The RoundUp Ready Story
Glyphosate is a broad-spectrum herbicide

Active ingredient in RoundUp herbicide
Kills all plants it come in contact with
Inhibits a key enzyme (EPSP synthase) in an amino acid pathway
Plants die because they lack the key amino acids
A resistant EPSP synthase gene allows crops

to survive spraying
RoundUp Sensitive Plants
+ Glyphosate
X
Plant
EPSP synthase
Without amino acids,

X
3-Enolpyruvyl shikimic acid-5-phosphate
(EPSP)
plant dies
X
X
Aromatic
amino acids
RoundUp Resistant Plants
+ Glyphosate
RoundUp has no effect;
Bacterial enzyme is resistant to herbicide
EPSP synthase
3-enolpyruvyl shikimic acid-5-phosphate

(EPSP)
With amino acids,
plant lives
Aromatic
amino acids
The Golden Rice Story
Vitamin A deficiency is a major health problem
Causes blindness
Influences severity of diarrhea, measles
>100 million children suffer from the problem
For many countries, the infrastructure doesnt exist

to deliver vitamin pills
Improved vitamin A content in widely consumed crops

an attractive alternative
-Carotene Pathway in Plants
IPP
Phytoene synthase
Phytoene
Problem: Phytoene desaturase
Rice lacks
these enzymes -carotene desaturase
Lycopene
Lycopene-beta-cyclase
Normal
Vitamin A -carotene
Deficient (vitamin A precursor)
Rice
The Golden Rice Solution
-Carotene Pathway Genes Added
IPP
Daffodil gene Phytoene synthase
Phytoene
Vitamin A
Phytoene desaturase
Pathway Single bacterial gene;
is complete performs both functions
and functional -carotene desaturase
Lycopene
Daffodil gene Lycopene-beta-cyclase
Golden -carotene
Rice (vitamin A precursor)
Transformation Cassettes
Contains
1. Gene of interest
The coding region and its controlling elements
2. Selectable marker
Distinguishes transformed/untransformed plants
3. Insertion sequences
Aids Agrobacterium insertion
Gene of Interest
Promoter TP Coding Region
Promoter Region
Controls when, where and how much the gene is expressed
ex.: CaMV35S (constitutive; on always)
Glutelin 1 (only in rice endosperm during seed development)
Transit Peptide
Targets protein to correct organelle
ex.: RbCS (RUBISCO small subunit; choloroplast target
Coding Region
Encodes protein product
ex.: EPSP
-carotene genes
Selectable Marker
Promoter Coding Region
Promoter Region
Normally constitutive
ex.: CaMV35s (Cauliflower Mosaic Virus 35S RNA promoter
Coding Region
Gene that breaks down a toxic compound;
non-transgenic plants die
ex.: nptII [kanamycin (bacterial antibiotic) resistance]
aphIV [hygromycin (bacterial antibiotic) resistance]
Bar [glufosinate (herbicide) resistance]
Effect of Selectable Marker
Non-transgenic = Lacks Kan or Bar Gene
Plant dies in presence

X
Transgenic = Has Kan or Bar Gene
Plant grows in presence

Insertion Sequences
TL TR
Required for proper gene insertions

Used for Agrobacterium-transformation
ex.: Right and Left borders of T-DNA
Lets Build A Complex Cassette
pB19hpc (Golden Rice Cassette)
TL aphIV 35S Gt1 psy 35S rbcS crtl TR
T-DNA Hygromycin Phytoene Phytoene T-DNA

Border Resistance Synthase Desaturase Border
Insertion Selectable Gene of Gene of Insertion

Sequence Marker Interest Interest Sequence
Delivering the Gene
to the Plant
Transformation cassettes are developed in the lab
They are then introduced into a plant
Two major delivery methods
Agrobacterium
Tissue culture
Gene Gun required to generate
transgenic plants
Plant Tissue Culture
A Requirement for Transgenic Development
Callus
grows
A plant part Shoots
Is cultured develop Shoots are rooted;
plant grows to maturity
Agrobacterium
A natural DNA delivery system
A plant pathogen found in nature

Infects many plant species
Delivers DNA that encodes for plant hormones
DNA incorporates into plant chromosome
Hormone genes expressed and galls form at infection site
Gall on
stem
Gall on
leaf
The Galls Can Be
Huge
The Lab Steps
Lab Testing The Transgenics
Insect Resistance Cold Tolerance
Bt-toxin protein CBF transcription factors
More Modern Examples
Salt Tolerant Mercury Resistance
Glyoxylase I Mercuric ion reductase
The Next Test Is The Field
Herbicide Resistance
Non-transgenics
Transgenics
Final Test
Consumer Acceptance
RoundUp Ready Corn
Before After
Infection of plants by Agrobacterium
Ti Plasmid of Agrobacterium
Transformation of Plants by Agrobacterium
Select for with antibiotic or herbicide resistance
cloning
Transgenic tobacco harboring a GFP Gene
Transgenic Cotton
Bt transformed cotton untransformed cotton

Transgenic herbicide resistant corn
European corn borer
Bt transformed corn is resistant to this insect

Biosynthetic Pathway for Vitamin A
(daffodil)
(bacteria)
Development of Golden Rice
Idea: Engineer rice to produce vitamin A
Vitamin A deficiencies primarily affect vision and leads to increased

susceptibility to infection and cancer
Vitamin A deficiency prominent in developing countries affects 124

million children worldwide; in SE Asia: 250,000 children go blind annually
See Also: http://www.uaf.edu/chem/451f00D/goldenrice/

WT and Transgenic Rice
WT golden rice
Microinjection vs particle bombardment
Reverse Genetics
ON/OFF Switch Makes Protein stop sign
PROMOTER INTRON CODING SEQUENCE poly A signal
Plant Transgene
Plant Selectable
Marker Gene
Plasmid DNA
Construct
bacterial genes
antibiotic marker
replication origin

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VISTAS

19th March, 2008

Paleolithic society Hunter-gatherers Nomadic

Long history of fermented foods since people

Dough not baked immediately would undergo

Uncooked fermented dough could be used to

1866 Louis Pasteur published his findings on

1915 Production of bakers yeast

Chemical transformations to produce therapeutic

Substrate + Microbial Enzyme Product

Microbial synthesis of other commercially valuable

Amino acids to improve food taste, quality or

Enzymes (cellulase, collagenase, diastase,

W.H. Sutton 1902

T.H. Morgan Drosophila melanogaster

Beadle and Tatum (Neurospora crassa)

Hershey and Chase 1952

Watson, Crick, Franklin and Wilkins (1953)

DNA Revolution Promise and Controversy!!!

Scientific foundation of modern biotechnology

1859 Charles Darwin publishes On

1864 Louis Pasteur shows all living

1865 The age of genetics begins

1902 Walter Sutton coins the term

1928 Fleming discovers antibiotic

1941 George Beadle and Edward Tatum propose

1949 Sickle cell anaemia demonstrated to be

1953 The double helix is

1967 The genetic code is cracked

1973 Recombinant DNA

1975 First international conference

1975 Monoclonal antibody

1978 Genentech Inc. established

1978 Genentech use genetic engineering to produce

1978 Kary Mullis discovers PCR

1996 First mammal cloned from adult

1990s First conviction using genetic

1996 Development of Affymetrix

1997 First artificial chromosome

An RNA Club organized by George Gamow (1954)

Vernon Ingrams research on sickle cell anemia (1956)

Radioactive tagging experiments demonstrate

Transgenic organism: an organism with artificially

Production of new and improved crops/foods,

The clone with her

Now researchers have

Carbohydrates Fermentation Carbohydrates

Desired Gene (crosses)

Poplar Soybean Ohio Buckeye

+ CP4 EPSPS = Roundup

Soybeans: Roundup Ready

Insect resistant cotton

Insect resistant corn

bST (bovine somatotropin)

SOIL EROSION INEFFICIENT

Benefits of biotechnology More food

Benefits of biotechnology Better food

1.Identify the desired

Crops are genetically improved to tolerate

targeting sequence (optional)

cauliflower mosaic virus 35S promoter

May lead to low genetic variability

These mice have been inserted with a jellyfish gene

National Geographic article about glowing mice