1. Recombinant DNA
Technology
PRESENTED BY:
D.PRIYANKA
M-PHARM
DEPARTMENT OF PHARMACEUTICS
UNDER GUIDENCE OF:
Mrs.YASMIN BEGUM
ASSOCIATE PROFFESSOR(Ph.D)
MALLA REDDY COLLEGE OF PHARMACY

2. CONTENTS:
Definition
RECOMBINANT dna TECHNOLOGY
RESTRICTION ENZYMES AND PLASMIDS
DEFINITION OF GENE
GENE CLONING
BASIC STEPS IN GENE CLONING
APPLICATIONS OF rdna technology
Conclusion
references

3. What is DNA?
DNA= Deoxyribu-Nucelic Acid
 DNA is a very large molecule,
made up of smaller units called
nucleotides
 Each nucleotide has three parts: a
sugar (ribose), a phosphate
molecule, and a nitrogenous base.
 The nitrogenous base is the part of
the nucleotide that carries genetic
information
 The bases found in DNA are four:
adenine, cytosine, guanine, and
thymine ( ATP, CTP, GTP, and
TTP)

4. Recombinant DNA Technology
 Recombinant DNA technology
procedures by which DNA from
different species can be isolated, cut
and spliced together -- new
"recombinant " molecules are then
multiplied in quantity in populations of
rapidly dividing cells (e.g. bacteria,
yeast).

5. Recombinant DNA Technology
 In the early 1970s it became possible to
isolate a specific piece of DNA out of the
millions of base pairs in a typical genome.

6. Recombinant DNA Technology
Recombinant DNA technology is based on a
number of important things:
 Bacteria contain extra chromosomal
molecules of DNA called plasmids which
are circular.

7. Recombinant DNA Technology
 Bacteria also produce enzymes called
restriction endonucleases that cut DNA
molecules at specific places into many smaller
fragments called restriction fragments.
 There are many different kinds of restriction
endonucleases

8. Recombinant DNA Technology
Restriction Enzymes and plasmid
 Sticky end and blunt end are the two
possible configurations resulting from the
breaking of double-stranded DNA

9. Recombinant DNA Technology
Restriction Enzymes and plasmid
 When RES acts at the center of symmetry, two
complementary strands of DNA are of equal
length, hence forms the blunt end.
T C A G A T C A GA
A G T C T A G T CT

10. Recombinant DNA Technology
Restriction Enzymes and plasmid
 Some RES breaks the DNA on either side of
center of symmetry with the liberation of
unequal fragments which are called as stick
ends/ cohesive ends.
 G A A T T C G A A T T C
 C T T A A G C T T A A G

11. Recombinant DNA Technology
Digestion of DNA by EcoRI to produce
cohesive ends.

12. Recombinant DNA Technology
Restriction Enzymes and plasmid
 Restriction Enzymes are primarily found in
bacteria and are given abbreviations based
on genus and species of the bacteria.
 One of the first restriction enzymes to be
isolated was from EcoRI
 EcoRI is so named because it was isolated
from Escherichia coli strain called RY13.

13. What is gene?
• A gene is a stretch of DNA
that codes for a type of
protein that has a function
in the organism.
• It is a unit of heredity in a
living organism.. All living
things depend on genes
• Genes hold the information
to build and maintain an
organism's cells and pass
genetic traits to offspring.

14. Gene cloning
 It can be defined as the isolation and
amplification of an individual gene
sequence by insertion of that individual
gene sequence into a bacterium where it
can be replicated

15. BASIC STEPS IN GENE CLONING
Step 1
A fragment of DNA,
containing the gene to
be cloned, is inserted
into a circular DNA
molecule called a
vector, to produce a
chimera or recombinant
DNA (rDNA)
molecule.
15

16. Step 2
The vector acts as a vehicle that transports the gene
into a host cell, which is usually a bacterium
although other types of living cell can be used. This
process is called transformation.

17. Step 3
Within the host cell the vector multiplies producing
numerous identical copies not only of itself but also
of the gene that it carries.

18. 18
Step 4
When the host cell
divides, copies of
rDNA molecule are
passed to the
progeny and further
vector replication
takes place.

19. Step 5
After large no: of cell divisions a colony or
clone of identical host cells is produced. Each
cell in the clone contains one or more copies of
the rDNA molecule
Step 6
Then, the host cells are then lysed and rDNA
can be separated.

20. Applications of rdna technology in medicine
Recombinant DNA technology had made it possible to treat
different diseases by inserting new genes in place of
damaged and diseased genes in the human body.
Insulin:-
Insulin is a hormone made up of protein. It is secreted in
the pancreas by some cells called as islet cells. If a
person has decreased amount of insulin in his body, he
will suffer from a disease called diabetes. Recombinant
DNA technology has allowed the scientists to develop
human insulin by using the bacteria as a host cell and it
is also available in the market. It is believed that the
drugs produced through microbes are safer.

21. VACCINES:
Recombinant DNA technology enables the
scientists to develop vaccines by cloning the gene used
for protective antigen protein. Viral vaccines are most
commonly developed through this technology for
example, Herpes, Influenza, Hepatitis and Foot and
Mouth Diseases
Human Growth Hormones:-
In recent years, scientists have developed many growth
hormones using recombinant DNA technology. The
disease of dwarfism is treated with this hormone.

22. Infectious Diseases:-
Many diseases are diagnosed by conducting certain tests.
Recombinant DNA technology has allowed the development
of many tests which are being used to diagnose diseases like
TB and cancer.
In the diagnosis process, certain pathogens are isolated and
identified, and then diagnostic kits are produced when the
genome of the specific pathogen is known to kill it or block
its pathogenic activity.

23. PRODUCTION OF NOVEL PLANTS:
Rdna is used in distinguishing of novel agricultural plants
which are high yielding and pest resistant
Cloning of genes from wild pest resistant varieties has been
used.
Strain improvement for fermentation:
Rdna uses extensively for improvement of strains of
microbes.

24. REFERENCES:
 FUNDAMENTALS OF MEDICAL
BIOTECHNOLOGY: Author: Aparna Raja
Gopalan,editors: irfan ali khan, page no:203-226
 U.Sathyanarayana: biotechnology: page no: 530-542
 pharmaceutical biotechnology: fundamentals and
applications: Author: s s kori. Page no:74-80.