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Transgenic Principles Seminar

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16 views12 pages

Transgenic Principles Seminar

Uploaded by

venkatesang
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© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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TRANSGENIC PRINCIPLES

IN PLANT AND ANIMAL

- URK20BT1021 ABIRAMI.R
What is meant by “Transgenic” ?
 Relating to or denoting an organism that contains genetic
material into which DNA from an unrelated organism has
been artificially introduced.
 Transgenic organisms are generated in the laboratory for
research purposes.

Principle:
 The basic principle of developing transgenic plants and
animals is to introduce the gene of interest into the
nucleus of germ cells, so that it can be passed onto
future generations as well.
Transgenic principle in animals
 The three principal methods used for the creation
of transgenic animals are
1. DNA microinjection
2. Embryonic stem cell-mediated gene transfer
3. Retrovirus-mediated gene transfer
 For practical reasons, i.e., their small size and low cost of housing in
comparison to that for larger vertebrates, their short generation time, and
their fairly well defined genetics, mice have become the main species used
in the field of transgenics.
 EX : Transgenic (Mice, rabbits , chicken, sheep,cow , donkey,monkey,fish)
1. DNA microinjection:
 It is one of the first methods that proved to be effective in mammals (Gordon and
Ruddle, 1981)
 This method involves the direct microinjection of a chosen gene construct (a single
gene or a combination of genes) from another member of the same species or from a
different species, into the pronucleus of a fertilized ovum.
 A major advantage of this method is its applicability to a wide variety of species.

2. Embryonic stem cell-mediated gene


transfer:
 This method involves prior insertion of the desired DNA sequence by homologous
recombination into an in vitro culture of embryonic stem (ES) cells.
 These cells are then incorporated into an embryo at the blastocyst stage of
development. The result is a chimeric animal.
 ES cell-mediated gene transfer is the method of choice for gene inactivation, the so-
called knock-out method.
 This technique is of particular importance for the study of the genetic control of
developmental processes.
 This technique works particularly well in mice. It has the advantage of allowing
precise targeting of defined mutations in the gene via homologous recombination.

3. Retrovirus-mediated gene transfer


 To increase the probability of expression, gene transfer is mediated by means of a
carrier or vector, generally a virus or a plasmid.
 Retroviruses are commonly used as vectors to transfer genetic material into the
cell, taking advantage of their ability to infect host cells in this way.
 Offspring derived from this method are chimeric, i.e., not all cells carry the
retrovirus.
 Transmission of the transgene is possible only if the retrovirus integrates into
some of the germ cells
TECHNIQUES:
 Physical Transfection
In this method, the gene of interest is directly injected into the pronucleus of a
fertilized ovum. It is the very first method that proved to be effective in mammals.
This method was applicable to a wide variety of species. Other methods of physical
transfection include particle bombardment, ultrasound and electroporation.
 Chemical Transfection

One of the chemical methods of gene transfection includes transformation. In this


method, the target DNA is taken up in the presence of calcium phosphate. The
DNA and calcium phosphate co-precipitates, which facilitates DNA uptake. The
mammalian cells possess the ability to take up foreign DNA from the culture
medium.
 Retrovirus-Mediated Gene Transfer

To increase the chances of expression, the gene is transferred by means of a


vector. Since retroviruses have the ability to infect the host cell, they are used as
vectors to transfect the gene of interest into the target genome.
 Viral Vectors
Viruses are used to transfect rDNA into the animal cell. The viruses possess the ability to
infect the host cell, express well and replicate efficiently.
 Bactofection
It is the process by which the gene of interest is transferred into the target gene with the
help of bacteria.

Applications of Transgenic Animals :


 Normal Physiology and Development
 Study of Diseases
 Study of Diseases
 Study of Diseases
Transgenic Plants :
 “A transgenic plant is a modified organism where genes are transferred from one
organism to another through genetic engineering techniques”.
 The purpose of producing a transgenic plant is to obtain a species that has ideal traits,
high yield and quality.

Methods Used for Gene Transfer:


The two methods include:
1. Agrobacterium mediates gene transfer
2. Particle bombardment / Gene gun method
Agrobacterium mediates gene transfer
Gene gun method
Examples of Transgenic Plants:
 Golden rice
 Bt cotton
 Flavr Savr

 Several other plants like potato, corn, papaya, maize squash, pumpkin, alfalfa, etc
have been modified to obtain crops with better yield and resistance.

Applications of Transgenic Plants:


 Resistance to biotic and abiotic stress
 Increased nutritional value
 Factories for production of recombinant proteins
THANK YOU

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