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BIOTECHNOLOGY

FOR THE YOUNG


Dr.
NEERAJA SOOD
MINDS
DYAL SINGH COLLEGE
UNIVERSITY OF DELHI
NEW DELHI

BIOTECHNOLOGY
The use of living organisms or their products
to enhance our lives and our environment.
A more narrow definition is;
The commercial application of living
organisms or their products, which involves
the deliberate manipulation of their DNA
molecules"

WHY BIOTECHNOLOGY?
WHY BIOTECHNOLOGY?
Traditional methods of improvement are
limited in scope due to genetic barriers
Improvement may not be of high standards
Methods of improvement are time
consuming.
Markers associated with improvement are
limited (diagnostic tests to identify genes
for inherited diseases like cystic fibrosis,
Huntingtons disease).

TECHNIQUES USED IN
BIOTECHNOLOGY

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r-DNA TECHNOLGY

TRANSgenesis

TRANSGENICS
A large number of transgenic animals
have been created
Mice Cows Pigs Sheep Goats Fish
Frogs Insects
Currently, no transgenic animal or
animal product is approved by the
FDA or USDA for human consumption

TRANSGENIC CATTLE
Dairy cows carrying extra copies of two types
of casein genes produce 13% more milk protein
Not only will this make the milk more nutritious,
it would allow for less milk to make more
cheese
Currently the milk from these animals is under
FDA review
The important difference between this & other
transgenics is that the DNA added is not
foreign

EnviroPig

TM

Transgenic pigs express phytase in their salivary


glands
Phytic acid in the pig meal is degraded releasing
phosphorus
The phosphorus is absorbed by the pig
Normally the phytic acid/phosphorus complex is
excreted
Pig waste causes eutrophication of lakes & streams

Transgenic Fish
Tilapia
Salmon/trout
Catfish
All can grow up to 6 times faster than
wildtype fish
Most have extra copies of growth hormone
(GH) gene

TRANSGENIC FISH
GloFish, originally developed in Singapore as a way to
monitor water pollution
The normally black-and-silver zebrafish was turned green
or red by inserting various versions of the GFP gene
Glofish are on sale throughout the US except in California
Glofish retail for about $5 per fish. Normal zebra-fish cost
around one tenth of the price

Transgenic Crops
Transgenic crops resistant to
insect pests
Transgenic crops resistant to
diseases
Drought tolerant transgenic crops
Herbicide tolerant transgenic
crops
Transgenics with nutritive traits
like Golden Rice

Insect Protective Genes


Bt cry genes from Bacillus
thuringiensis
highly effective, highly specific, safe
to environment and higher animals,
amenable to genetic engg.
Trypsin Inhibitor genes
Lectin genes

DNA ENGINEERING

VACCINES
(EDIBLE)

IMMOBILISATION OF ENZYMES

LACONES

CONSERVATION CLONING
Many endangered or extinct animals are
being cloned or considered for cloning
Gaur
Bucardo mountain goat
Mammoth
Quagga
Banteng

CLONING

Dolly, First Mammal Cloned From an Adult Cell

Dolly, as an adult

GENE CLONING
IDENTIFICATION OF DESIRABLE GENES (cut DNA
by RE -> gel electrophoresis -> elution of
band/desirable DNA)
INTRODUCTION OF IDENTIFIED DNA INTO THE
HOST(use of cloning
vectors/RE/ligases/transformation )
MAINTAINENCE OF INTRODUCED DNA IN THE
HOST AND ITS TRANSFER IN PROGENY(use of
selectable marker)
OBTAINING THE RECOMBINANT PROTEIN
(bioreactor)

pBR322

BACTERIOPHAGE LAMDA

CLONING VECTOR

VECTORS
Fig 2. YAC

Fig 1.BAC

RE

r-Plasmid DNA

COLONY HYBRIDISATION

PCR

HUMAN INSULIN PRODUCTION BY


BACTERIA

FERMENTORS

Mammalian Cell Bioreactor

DOWNSTREAM PROCESSING

Bacterial and Animal Biotechnology Products


Biotech chymosin
enzyme used to curdle milk products
gene from yeast
harvested from GE bacteria
replaces the calf enzyme
bST (bovine somatotropin)
increases milk production
gene from cow
protein harvested from GE bacteria
replaces cow protein originally

harvested from pituitary glands

of slaughtered cows

SCOPE OF BIOTECHNOLOGY

Human Health
Diagnostics (e.g., biosensors, immunodiagnostics, gene
probes)
Therapeutics (e.g., vaccines, immune stimulants,
biopharmaceuticals)
Drug Delivery
Bioinformatics
Genomics and Molecular Modelling (e.g., DNA/RNA/protein
synthesizing and databases for humans, plants, animals, and
micro-organisms)
Gene Therapy (e.g., gene identification, gene constructs, gene
delivery)

SOME MORE AREAS


Environment
Air, Water and Soil (e.g., bioremediation,
diagnostics,
phytoremediation,biofiltration)
Natural Resources
Energy (e.g., microbiologically enhanced
petroleum recovery,
industrial bioprocessing, biodesulphurization)
Mining (e.g. microbiologically enhanced mineral
recovery,
industrial bioprocessing, biodesulphurization)
Forest Products (e.g., biopulping, biobleaching,
biopesticides,
tree biotechnology, industrial bioprocessing)

ADVANCES: In 1977, Genentech reports production of first


human protein, Somatostatin, in bacteria
Genentech Inc and the City of Hope National
Medical Center announces production of human
insulin using recombinant technology
US supreme court allowed patenting of
genetically altered life; oil eating microbe
patented
GE plants are also patented
USDA approves release of GE tobacco plants
under field conditions

Recombinant hepatitis B vaccine approved


Centocars CA125TM test for ovarian cancer
approved
Genentech Inc. gets approval to market rtPA (GE tissue plasminogen activator) to
treat heart attacks
Anticancer drugs, Interleukin-2 and
polyethylene glycol IL-2 licensed
Kary Mullis introduced PCR in 1988
The Human Genome Project, the
international effort to map all of the genes
in the human body, was launched.
Estimated cost: $13 billion. 1990 Formal
launch of the international Human Genome
Project

Southern Blotting

VNTRs
Every strand of DNA has pieces that contain genetic
information which informs an organism's development
(exons) and pieces that, apparently, supply no relevant
genetic information at all (introns). Although the introns
may seem useless, it has been found that they contain
repeated sequences of base pairs. These sequences,
called Variable Number Tandem Repeats (VNTRs), can
contain anywhere from twenty to one hundred base pairs.
Every human being has some VNTRs. To determine if a
person has a particular VNTR, a Southern Blot is
performed, and then the Southern Blot is probed, through
a hybridization reaction, with a radioactive version of the
VNTR in question. The pattern which results from this
process is what is often referred to as a DNA fingerprint.
A given person's VNTRs come from the genetic
information donated by his or her parents; he or she
could have VNTRs inherited from his or her mother or
father, or a combination, but never a VNTR either of his or
her parents do not have. Shown below are the VNTR
patterns for Mrs. Nguyen [blue], Mr. Nguyen [yellow], and
their four children: D1 (the Nguyens' biological daughter),
D2 (Mr. Nguyen's step-daughter, child of Mrs. Nguyen and
her former husband [red]), S1 (the Nguyens' biological
son), and S2 (the Nguyens' adopted son, not biologically
related [his parents are light and dark green]).

VNTR

DNA FINGERPRINTING

The chemical structure of


everyone's DNA is the same.
The only difference between
people (or any animal) is the
order of the base pairs.
There are so many millions
of base pairs in each
person's DNA that every
person has a different
sequence.
Using these sequences,
every person could be
identified solely by the
sequence of their base
pairs. However, because
there are so many millions
of base pairs, the task
would be very timeconsuming. Instead,
scientists are able to use a
shorter method, because of
repeating patterns in DNA.
These patterns do not,
however, give an individual
"fingerprint," but they are
able to determine whether
two DNA samples are from

5' T-T-G-A-C-T-A-T-C-C-A-GA-T-C 3'


3' A-A-C-T-G-A-T-A-G-G-T-CT-A-G 5'

The chemical structure of DNA is as follows:

DNA FINGERPRINTING

Different individuals carry


different
alleles.
Most alleles
useful for DNA fingerprinting differ on the
basis of the number of repetitive DNA sequences they
contain.

HGP
Human Genome Project has been called a Mega Project
because of the following factors:
1. The human genome has approx. 3.3 billion base-pairs; if
the cost of sequencing is US $3 per base-pair, then the
approx. cost will be US $10 billion.
2. If the sequence obtained were to be stored in a typed form
in books and if each page contains 1000 letters and each
book contains 1000 pages, then 3300 such books would be
needed to store the complete information.
However, if expressed in computer storage units (3.3 billion
base-pairs) x (2 bits per pair) = 825 megabytes of raw data.
Which is about the same size of one music CD. If further
compressed, this data can be expected to fit in less than 20
Megabytes.

CENTRAL DOGMA

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NEXT STEP?

PLAYING GOD!
Synthetic Mycoides
refers to an artificial life
form created by
Craig Venter at the
J Craig Venter Institute in
May 2010. A synthetic
genome was transferred
into an empty cell to
form the bacterium.

J. CRAIG VENTOR

THANKS!