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PRINCIPLES, PROCESSES
AND
APPLICATIONS
BY
B.H.SOORYA RAO
CLASS XII
DEFINITION
1.
2. GENETIC
CULTURE
The
It
facilitates
technique
ENGINEERING
TECHNIQUES
theofgrowth
altering
and
chemistry
multiplication
of genetic
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the desired
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m
PRINCIPLES
OF
MODERN
BIOTECHNOLOGY
WHAT IS CLONING?
Cloning or making multiple copies of
any template DNA needs that an alien
DNA is linked with the origin of
replication (a desired DNA sequence
responsible for initiating replication) so
that alien (foreign/desired) pieces of
DNA can replicate and multiply itself in
the host organism.
RESTRICTION ENZYMES
When cut by the same RE, the resultant DNA fragments have the same
kind of sticky ends and these can be joined together using DNA ligase.
NOMENCLATURE OF RESTRICTION
ENZYMES
The first letter of the name comes from genus and the next two
letters from the name of the species of the species of the bacterium
(prokaryotic cell) from which they are isolated.
The Roman number following these four letters indicate the order in
which enzymes were isolated from that strain of the bacterium, e.g.
(a) Eco R-I is isolated from Escherichia coli RY 13
(b) Hind- II is from Haemophilus influenzae
(c) Bam H-I is from Bacillus amyloliquefaciens
(d) Sal-II is from Streptomyces albus.
CLONING VECTOR
CLONING VECTOR
FEATURES OF VECTOR
Selectable marker : It helps in identifying ang eliminating nontransformants and help in selecting those host cells which contain
the vector, i.e., transformants.
PROCESSES OF
RECOMBINANT DNA
TECHNOLOGY
IN DETAIL
1.
ISOLATION OF DNA
DNA is enclosed by membranes along with other macromolecules such as
RNA, proteins polysaccharides and lipids.
(a) The membrane is dissolved by treating the bacterial cells/plant or animal
tissue with enzymes such as lysozyme (bacteria), cellulase (plant cell),
chitinase (fungus).
(b) Long DNA molecules are interwined with histone proteins. RNA can be
removed by treating with ribonuclease whereas the proteins can be removed
by proteases.
(c) Other molecules can be removed by treating by appropriate treatments
and now purified DNA precipitates out after the addition of chilled ethanol.
(d) It is seen as collection of fine thread in the suspension.
2.
3. SEPARATION AND
ISOLATION OF DNA
FRAGMENTS
5.
JOINING OF DNAs
(a) After cutting DNA with specific RE, the cut out gene of interest from the
source DNA and the cut vector with space are mixed and ligase is added.
(b) This results in the preparation of recombinant DNA.
6.
RECOMBINANT PROTEIN
Recombinant protein is produced if any protein encoding
gene is expressed in heterologous host.
(i) The transformed cells containing cloned gene of interest
are grown in cultures
BIOREACTORS
Bioreactors are used for processing large volume of
culture for obtaining the product of interest in large
quantities. In bioreactors the raw materials are
converted into specific products, e.g., enzymes.
A bioreactor provides the optimal conditions for
achieving the desired product by providing optimal
growth condition such as:
(i) temperature
(ii) pH
(iii) substrate
(iv) salt
(v) vitamins
(vi) oxygen
BIOREACTORS
APPLICATIONS OF
BIOTECHNOLOGY
REASEARCH AREAS IN
BIOTECHNOLOGY
Three critical research areas of biotechnology are :
BIOTECHNOLOGICAL APPLICATIONS IN
AGRICULTURE
Three options for increasing food production :
1. Agrochemical based agriculture
2. Organic agriculture
3. Genetically engineered crop based agriculture.
2.
A novel strategy was adopted to prevent this infestation which was based
on the process of RNA interference (RNAi). RNAi takes place in all
eukaryotic organisms as a method of cellular defense. This method
involves silencing of a specific mRNA due to a complementary dsRNA
molecule that binds to and prevents translation of the mRNA (silencing).
3.
4.
5.
The introduction of DNA was such that it produced both sense and antisense RNA in the host cells.
6.
7.
The consequence was that the parasite could not survive in a transgenic
host expressing specific interfering RNA.
8.
The transgenic plant therefore got itself protected from the parasite
GENE THERAPY
MOLECULAR DIAGNOSIS
Recombinant DNA Technology, polymerase chain reaction
(PCR), and Enzyme-linked Immunosorbent Assay (ELISA) are
some of techniques diagnosis because :
1. Very low concentration of bacteria or virus (before appearance of visible
symptoms of disease) can be detected by amplification of their nucleic acids.
2. PCR is now used to detect HIV in suspected AIDS patients.
3. A probe is a piece of single stranded DNA that is tagged with a radioactive
molecule and it is used to find it complementary by hybridization.
4. Presence of normal or mutated gene can be detected by this method .
TRANSGENIC ANIMALS
Transgenic animals are those that have had their DNA manipulated to
posses and express an extra or foreign gene, example transgenic rats, rabbits,
pigs, sheep, cow, etc.
ETHICAL ISSUES
BIOPIRACY
Biopiracy is the term used to refer to the use of bioresources
by MNCs and other organizations without proper authorization
from the country and people concerned without compensatory
payment.
(i) Financially rich industrialized nations but poor in biodiversity
exploit the bioresources and traditional knowledge of developing and
underdeveloped poor countries rich in biodiversity.
(ii) Some nations are developing laws to prevent such unauthorized
exploitation of their bioresources and traditional knowledge.