Recombinant DNA Tech

1. RECOMBINANT DNA
TECHNOLOGY
SUBMITTED BY:- ROLL NO 56-60
DVM 3RD SEMESTER
SESSION 2019-2024
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2. Definition:-
“Recombinant DNA Technology involves the
artificial synthesis, modification, removal, addition and repair of
DNA”
Recombinant DNA Technology is a beneficiary branch of
“GENETIC ENGINEERING”
It is used in cases of end stage diseases, manufacturing
processes of hormones for medical uses etc.
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3. History of Recombinant DNA Technology:-
 Discovered by Herbert W Boyer and Stanley Norman Cohen
in 1973.
 Paul Berg created first Recombinant DNA molecule at
Stanford.
 Humulin was the first drug produced via R DNA tech in 1982
and this was an insulin.
 FDA approved Humulin in October 1982.
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4. Objectives of Genetic Engineering:-
1. For Gene Therapy as the final resource of cure.
2. Production of drugs, organic chemicals and enzymes.
3. For proteins and hormone production e.g. Insulin.
4. Production of better varieties of plants that are pest and
disease resistant.
5. Large scale production of animal based products in lesser
time.
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5. Steps of Recombinant DNA Technology:-
The basic steps in any Genetic Engineering Technique remain the
same while distinguishing steps are different. Basic steps are;
1. Isolation of gene of interest.
2. Insertion of gene of interest to a vector.
3. Transfer of Recombinant DNA into host organism.
4. Growth of genetically modified microbe.
5. Expression of gene.
6. Isolation of desired chemicals or hormones.
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6. Explanation:-
1. Isolation of Gene of Interest:-
In this first step, the gene
which has coding sequence that produces the desired product is
extracted from the DNA. For this Restriction Endonucleases are
used which cut the DNA at a specific point and result in a base
sequence required.
Each gene has a specific Restriction Endonuclease for its cutting.
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7. 2. Insertion of Gene into Vector:-
The isolated gene is
now inserted into vector.
“Vector is the organism of organelle of microbe with which gene
of interest is attached”
This vector can be a bacteriophage virus, plasmid or DNA.
First Restriction Endonucleases are used to make place for
gene of interest in the plasmid and then Ligase will be used to
bind gene of interest into plasmid.
The vector will now be called Recombinant DNA.
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Sequential DNA ligation of gene of interest and insertion to vector.

9. 3. Transfer of Recombinant DNA into Host:-
The
Recombinant DNA is transferred to host by process called
“Micro-Injection.”
This host will now be called a GMO (Genetically Modified
Organism).
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10. Microinjection is the injection of
Something to an exceptionally small
Or microscopic sized object.
It uses very fine injection needles
and pipettes.
Microinjection to Cell
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11. 4. Growth of GMO:-
The GMO is now placed In Culture
Media for quick growth and multiplication. The culture media
has all the necessary nutrients required for optimal growth of
GMO.
This culture media can be NUTRIENT AGAR, GELATIN,
NUTRIENT BROTH etc.
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12. 5. Expression of Gene:-
The inserted gene of interest will
express itself and desire product will start to be synthesized.
This product is either released into the culture media or is
present in the organism as vesicles/vacuoles.
Collection is done by centrifuging the contents and separating
product from undesirable components.
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13. 6. Purification of Product:-
The final product is purified
through filtration techniques. This may be Chemical
filtration or filtration via filter paper/crucible.
The product may now be sterilized and packed to be distributed.
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14. Recombinant DNA Tech being used for growth hormone production.
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15. Where is Recombinant DNA Tech being used?
 In medical field, for therapies e.g. Gene Therapy or Bone Marrow
Transplant.
 In pharmaceutics, for manufacture of hormones and drugs e.g.
Insulin production, Interferon production.
 In plant breeding and genetics, for production of better Cultivars
and Varieties.
 in food industry, for solving problem of meat consumption. Here
Single cell protein and Lab grown meat technology are being
used.
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16. Commercial production of insulin in huge vats.
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17. Prevalent Risks in Genome Engineering:-
1. Always a risk of creating a mutant specie of microbes which
can cause Biohazards.
2. Minor faults in genome editing can lead to Faulty products.
3. Work on disease resistance can negatively lead to a New
Strain.
4. Terrorist organizations may reverse engineer a research and
cause Global Green Terrorism or Eco-Terrorism.
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18. Global Acceptability of Procedure:-
 In the beginning, this procedure was not being accepted by many
scientists an also Medical organizations but as the research went by
and discoveries happened. This technology was hailed as a success.
 People living in Rural areas who are illiterate don’t even accept
these procedure till now.
 Almost all of the global demands of Pituitary Hormones and Insulin
are met by the application of this technology.
 More and more research and development means this technology is
being accepted with a vast coverage.
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