Genetic
Engineering
Deals with the modification of genetic
material of an organism
4 Steps involved in
Genetic Engineering:
1.
01 DNA Cleavage
First we need to identify the genes or genes
that we are interested in, the target DNA.
Once the target and vector DNA have
been identified, both types of DNA are cut
using restriction endonucleases.
2.
02 Production of Recombinant DNA
Combine target and vector DNA – after both types
of DNA have been cleaved by the same restriction
endonuclease, the two types of DNA are
combined together with the addition of DNA
ligase, an enzyme that repairs the covalent bonds
on the sugar-phosphate backbone of the DNA.
3.
03 Cloning
Introduce recombined molecule into host cell –
once the target DNA has been stably combined
with vector DNA, the recombinant DNA must be
introduced into a host cell, in order for the genes Methods in inserting
to be replicated or expressed.
plasmid into Host Organism
1 Biolistics
A gene gun uses very small metal
particles (microprojectiles) coated with
the recombinant DNA, which are blasted
at a plant tissue at a high velocity. If the
DNA is transformed or taken up by the
cell’s DNA, the genes are expressed.
4.
04 Screening
It must go through a screening procedure
using various techniques to ensure that
the right host is chosen if it contains the
gene of interest.
Heat Shock Treatment 2 Methods to screen
The bacterial cells are treated with calcium
chloride and then suddenly exposed to high
temperatures. Heat shock is used to
temporarily form pores in the cell membrane,
allowing transfer of the exogenous DNA into
the cell.
3 PCR (Polymerase Chain Reaction)
Recombinant DNA
Polymerase chain reaction (abbreviated
1 Selection of plasmid DNA containing cells PCR) is a laboratory technique for rapidly
producing (amplifying) millions to billions of
A selection marker within the inserted copies of a specific segment of DNA, which
plasmid DNA sequence allows the can then be studied in greater detail.
selection of "transformants". Usually, an
antibiotic resistance gene is included in
the plasmid DNA. This mechanism allows
only "transformed" cells to survive in the
presence of aortic.

3 Electroporation
Why is there a need to modify the traits or
Selection of transformed cells with characteristics on a certain organism?
In this technique, cells are subjected to an
the desired gene (Hybridization)
2
electric field to make the bacterial cell The use of genetically modified organisms
The cloned genes form base-pairs with (GMOs) benefits both producers and
temporarily permeable.
complementary sequences on another nucleic
consumers in specific ways. Modified plants,
acid. The complementary nucleic acid is called
for instance, can at least initially help protect
as probe because it is used to probe for the
crops by providing resistance to a specific
presence of the gene of interest.
disease or insect, ensuring greater food
production. GMOs are significant sources of
medicine as well.

Alliyah Shay B. Muaña

XI Edison