Recombinant DNA

Joannah Mae C. Calico

MA.Ed _Biological Science
Definition of recombinant DNA
(rDNA)
-Combining two or more strands of DNA
(commonly DNA of two different
organism.
-DNA molecules are constructed in vitro,
then propagated in a host cell or
organism..
History of recombinant DNA
technology
Recombinant DNA technology is one
of the recent advances in
biotechnology, which was developed
by two scientists named Herbert
Boyer and Stanley N.Cohen in
1973.
Basic principle of recombinant
DNA technology

The DNA is inserted into another

DNA molecule called ‘vector’

. The recombinant vector is then

introduced into a host cell where it
replicates itself, the gene is then
produced
How is rDNA made?
Six steps of Recombinant DNA
1. Isolating
2. Cleavage/ Cutting
3. Ligation/ Joining
4. Transforming / Transfection
5. Cloning
6. Selecting
1. Isolation and Purification of
DNA
Both vector and target DNA molecules
can be prepared by variety of routine
methods (ultracentrifigation).

In some cases, target DNA is

synthesized in vitro. (PCR).
 2. Cleavage of DNA at
particular sequence
Cleaving DNA to generate fragment of
defined length or with specific
endpoints.

Insert DNA is produced, by treating it

with enzyme (restriction
endonuclease)
Cleavage of DNA
3. Ligation of DNA fragments
A recombinant DNA molecule is formed
by cleaving the DNA of interest to yield
insert DNA and then ligating it to the
vector DNA using DNA ligase.

Recombinant DNA= chimeric DNA

4. Introduction of rDNA into
compatible host cells.
In order to propagated , rDNA must be
introduced into a host cell where it can
replicate.
Transformation
-direct uptake of foreign DNA by host
cell (bacteria)
Transfection
-rDNA are packaged into virus particles
5. Replication and Expression
of rDNA in host cells.
Cloning vectors allow insert DNA to be
replicated and also expressed in a host
cell.

Efficiency of cloning depends on choice

of vectors and hosts.
 6. Identification of host cells
that contain rDNA of interest.

Vectors contain easily scored genetic

marker, which allow the selection of
host cells that have taken up foreign
DNA.
 Applications of
Recombinant DNA
Technology
Large-scale production of human
proteins by genetically
engineered bacteria.

Such as : insulin, Growth

hormone, Interferons and
Blood clotting factors (VIII & IX)
 Better Crops
Herbicide and pesticide resistance corn

“Golden rice” with beta-carotene

Recombinant Vaccines

Recombinant Pharmaceuticals
Transgenic Plants and Animals
 Summary
1. Recombinant DNA technology builds
on few basic techniques: isolation,
cleavage, ligation, introduction of DNA
into host cells and identification of cell
containing recombinants.
2. DNA fragment generated can be
ligated into cloning vectors including:
plasmids, bacteriophage, viruses and
artificial chromosomes.
3. Recombinant DNA methods allow the
production of proteins for therapeutic
use and identification of individual with
gene defects.
Thank you for listening!!!
Happy Dinagyang Festival.