IN
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
Examples:
Cholesterol Steroids (cortisone, estrogen,
progesterone) (hydroxylation reaction -OH
group added to cholesterol ring)
Classical biotechnology
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
Molecular Biology
Organismic biotechnology
uses intact organisms and does not alter genetic
material
Molecular Biotechnology
alters genetic makeup to achieve specific goals
Medicine
human
veterinary
biopharming
Environment
Agriculture
Food products
Industry and manufacturing
What are the applications of biotechnology?
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;
Grain Stover
Chemical
Pretreatment
Endosperm Germ
Containing Cellulose
Currently used suitable for
For ethanol production Cellulases
enzyme
degradation
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 =
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
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
Triticale
New species, but
NOT biotechnology
products
Agriculture Products On the Market
Source: USDA
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
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
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.
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.).
Novel gene
construct:
strong promoter
(sockeye MT) +
extra growth
hormone gene
(sockeye GH).
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
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
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%*
*= 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?
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 (%)
Brinjal
Mahyco, Mumbai cry1Ac
Sungro Seeds Ltd, New Delhi cry1Ac
IARI, New Delhi cry1F
Cabbage
Sungro Seeds Ltd, New Delhi cry1Ac
Cauliflower
Sungro Seeds Ltd, New Delhi cry1Ac
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
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
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
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.
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
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
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.
Biomedical Devices
Biomedical Instrumentation
Biosensors
Nanobiotechnology
Lessons:
Counter-examples:
High tech medicine
God wants us to
genetically engineer
food
Intrinsic objections to ag biotech
Counter-examples:
GIFT and in vitro
Plant cuttings
Intrinsic objections to ag biotech
Counter-example:
Agriculture
Intrinsic objections to ag biotech
4. Dont cross
species
Mules
Wheat
Intrinsic objections to ag biotech
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
New:
- spaghetti
- whisky
- ...
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
Aspartame
Artificial sweetener
Made from aspartic acid
and phenylalanine
Used in 5,000 products
Monosodium glutamate
Recombinant alpha amylase
Counter-intuitive result:
(Comstock slide)
Medicine v. Food
Positive attitudes toward
biomedical uses
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
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
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
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%)
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%.
$1500m
Indian Biotech Products & Services:
$ 150
$150m $ 100
$ 350
2002
2005
The Indian Biotech business stands at US$150 m today... 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
#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
#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
#19
R&D ORGAINZATIONS
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.
GROWTH 20%
RATE
Healthcare
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
#4
The India Advantage
Excellent network
Well-developed
of research laboratories
base industries
320
Opportunity in various Industry Sectors
Software
Biotechnology
Outsourcing
Retailing
Real Estate
Auto Components
Telecommunication
321
Opportunities in Biotechnology
322
Biotech Market - A consistent Uptrend
25 % growth in investment
70 % growth in employment
2002- 03
74 % growth in R&D
manpower
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
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biotechnology ...
biotechnology software, biotechnology consulting jobs.
India, the darling of the
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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.
...
www.bii.in/page/careers/job_career.asp
CareerAge - Career Choice - Biotechnology
India's complete Jobs & Career .companion ...
Biotechnology is an interdisciplinary
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subjects like mathematics, ...
www.careerage.com/resources/career_choices/bi
otech/biotech.shtml
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.
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
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
Researcher
$ $ 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
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:
Smooth Wrinkled
Allele
Tall Short
Allele
This Implies a
Genetic Continuum
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
is condensed
into
Chromosomes
Chromosome
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
3 5
5 3
5 3
5 3
PCR Animation
Clones transferred
to filter
Human clone
library PCR fragment
probe added to filter
Gene/Marker
2. Find cosegregating marker
Gene/Marker
4. Find ORFs on contig
Function:
Glyphosate (RoundUp) resistance
EPSP synthase enzyme
Increased Vitamin A content
Vitamin A biosynthetic pathway enzymes
The RoundUp Ready Story
+ Glyphosate
X
Plant
EPSP synthase
X
X
Aromatic
amino acids
RoundUp Resistant Plants
Shikimic acid + Phosphoenol pyruvate
+ Glyphosate
RoundUp has no effect;
Bacterial enzyme is resistant to herbicide
EPSP synthase
Aromatic
amino acids
The Golden Rice Story
Vitamin A deficiency is a major health problem
Causes blindness
Influences severity of diarrhea, measles
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
Geranylgeranyl diphosphate
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
Steps
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
TL TR
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
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
Introduce DNA
Agrobacterium or gene gun
Transgene= Transgene=
Bt-toxin protein CBF transcription factors
More Modern Examples
Salt Tolerant Mercury Resistance
Transgene= Transgene=
Glyoxylase I Mercuric ion reductase
The Next Test Is The Field
Herbicide Resistance
Non-transgenics
Transgenics
Final Test
Consumer Acceptance
Before After
The Public Controversy
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
Plant Transformation
Event Selection
Plant Breeding
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:
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
GMOs are an innovation that have and will continue to impact all
facets of the global agricultural economy.
Production Processing
2. Global trade
is condensed
into
Chromosomes
Chromosome
Gene
Homology Cloning
Clones transferred
to filter
Human clone
Mouse probe
library
added to filter
3 5
5 3
5 3
5 3
Gene Manipulation
Function:
Glyphosate (RoundUp) resistance
EPSP synthase enzyme
Increased Vitamin A content
Vitamin A biosynthetic pathway enzymes
PCR Animation
Clones transferred
to filter
Human clone
library PCR fragment
probe added to filter
Gene/Marker
2. Find cosegregating marker
Gene/Marker
4. Find ORFs on contig
+ Glyphosate
X
Plant
EPSP synthase
X
X
Aromatic
amino acids
RoundUp Resistant Plants
Shikimic acid + Phosphoenol pyruvate
+ Glyphosate
RoundUp has no effect;
Bacterial enzyme is resistant to herbicide
EPSP synthase
Aromatic
amino acids
The Golden Rice Story
Vitamin A deficiency is a major health problem
Causes blindness
Influences severity of diarrhea, measles
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
Geranylgeranyl diphosphate
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
TL TR
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
Gall on
stem
Gall on
leaf
The Galls Can Be
Huge
The Lab Steps
Lab Testing The Transgenics
Transgene= Transgene=
Bt-toxin protein CBF transcription factors
More Modern Examples
Salt Tolerant Mercury Resistance
Transgene= Transgene=
Glyoxylase I Mercuric ion reductase
The Next Test Is The Field
Herbicide Resistance
Non-transgenics
Transgenics
Final Test
Consumer Acceptance
Before After
Infection of plants by Agrobacterium
Ti Plasmid of Agrobacterium
Transformation of Plants by Agrobacterium
cloning
Transgenic tobacco harboring a GFP Gene
Transgenic Cotton
(daffodil)
(bacteria)
Development of Golden Rice
Idea: Engineer rice to produce vitamin A
WT golden rice
Microinjection vs particle bombardment
Reverse Genetics
Building the Transgenes
Plant Transgene
Plant Selectable
Marker Gene
Plasmid DNA
Construct
bacterial genes
antibiotic marker
replication origin