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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 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
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, 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
FERMENTORS
DOWNSTREAM PROCESSING
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)
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
DNA FINGERPRINTING
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!