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1. APPLICATION OF RECOMBINANT
DNA TECHNOLOGY
By: Misha Aanand
M.pharm Pharmacology sem 1
Cellular & molecular pharmacology(MPL104T)
Nirma University

2. What is recombinant technology?
Encompasses a number of experimental protocols leading to the transfer of genetic information(DNA) from
one organism to another. Involves the manipulation of genetic material (DNA) to achieve the desired goal in
a predetermined way.
Basic principles of rDNA technology:
● Generation of DNA fragments & selection of the desired piece of DNA.
● Insertion of the selected DNA into a cloning vector to create a rDNA or chimeric DNA.
● Introduction of the recombinant vectors into host cells.
● Multiplication & selection of clones containing the recombinant molecules.
● Expression of the gene to produce the desired product.

3. How recombinant and DNA came to exist?
Recombinant DNA technology is one of the recent advances in
biotechnology in which it became possible to isolate a specific
piece of DNA out of million of base pair in a typical genome
near 1970s.
Was first developed by 2 scientist name Boyer and Cohen in 1973.

4. Types of Recombinant technology
Majorly recombinant is done by 3 methods
Select a piece of DNA of interest
Cut DNA with
restriction enzyme
Ligate DNA with
vector by DNA ligase
Vector is inserted
2. Phage Introduction
1. Transformation
● Is same as Transformation, but here Phage ( Virus parasitic to bacteria) is used instead of Bacteria.
Phages: like MI3 and lambda.
3. Non- Bacterial transformation
● Microinjection, the DNA is injected directly into the nucleus of the cell being transformed. The host cells are
bombarded with high velocity microprojectiles, such as particles of gold or tungsten that have been coated
with DNA.

5. General application of DNA recombinant technology
● Application in Agriculture
-Increase Tolerance plant- Insect tolerated, Disease tolerated, Herbicidal tolerate and Stress tolerated
-High Quality and yielding plants
-Biopharmaceutical needed plant.
● Application in Environment
-Remedies of polluent
-Energy Formation- Cyanobacteria for CO2 to biofuel conversion, BOD during water sewage
management.
● Application in Industry
-Food industry. -Dairy Industry
- Pharmaceutical industry- Insulin, HGH, MoAB, Interferon
● Application in Forensic science
-DNA Fingerprinting- Hybridisation, PCR
- Sample analysis.
● Application in Medicine
-Vaccine preparation
-Diagnosis
-Gene therapy
-Antibodies

6. Application in agriculture
Plants have been modified since the last two decades by using conventional breeding methods but now through
transformation, recombinant DNA technology transgenic crops are being commercially produced.
Example.
Majorly it produces three types
1. Production of stress tolerant crops.
A) Herbicide tolerant plants
Many of time plant are killed due to grow of herbicides near those plants example round of herbicide which grow
near a plant and block EPSP synthase and kill the plant, so plant breeder take gene from those plants which have
resistant to word this herbicide and manipulate to insert in herbicide tolerant gene.
Soybean and cotton are modified EPSP gene crops
A) Insect tolerant plants
Before this technology pesticides and insecticides were used to kill harmful insects but were not eco friendly and
had adverse effect on crop as well as environment,
Best example is BT cotton, maize, rice, cabbage, cauliflower and many other crops BT gene is obtained from
Bacillus thuringiensis, is full for cry protein which forms toxin which binds to intestinal wall of insects who eat
the leaves of the plant.

7. C) Stress tolerant plant.
Menu of time due to unfavourable condition of environment like high temperature, acidic, soil salinity, high
amount of any ions present in soil cause stress on plant and leads to death but with the help of manipulation and r
DNA it can be modified two tall rate abiotic stress.
D) Disease resistance plants.
To save plants from various diseases caused by bacterial, viral or fungal infection, recombinant DNA technology
is used by inserting suitable vectors to form a defence mechanism against disease.
Example tobacco plant is modified by inserting for in genes to protect it from tobacco mosaic virus.
2. To increase quality and quantity of crops
To increase the yield of crops, photosynthetic genes are introduced in plants which leads to storage of starch in
tubers and not in leaves which leads to high yield.
Examples are
Administrating antisense RNA in tomatoes to slow down ripening and increase quality.
Ethylene is a hormone responsible for fruit ripening with the help of r DNA technology ethylene inducing genes
can be inserted to delay fruit ripening.
Vitamin A deficiency can also be treated by increasing production of beta carotene.
3. Production of biopharmaceuticals.
A) Edible Vaccine
B) Production of secondary metabolites.

8. Application in Environment
By recombinant DNA technology we can remediate environmental pollutants
● Some harmful chemicals that are not digested and degraded. They are partially degradable e.g. TNT
chemical. In this nitrogen is reacted with oxygen and superoxide is formed that is toxic.
knock out the monodehydroascorbate reductase gene that will create resistance against TNT
● Arsenic particles are toxic and they are present in the soil that is harmful for plants.
Pv ACR3 gene is inserted in Arabidopsis,
Pv ACR3 gene expression will generate a tolerance plant to arsenic.
In Arabidopsis thaliana reductase enzyme is present→ Reduce the arsenic
● By Recombinant DNA technology, plastic degradation can be enhanced by genetically modified
organisms.
● Degrade oil spills or organic waste.
Genetically modified strain of Pseudomonas putida able to degrade chemicals in oil spills.
● Used in development of bioindicators: bacteria have been genetically modified as 'bioluminescors' that
give off light in response to several chemical pollutants.

9. Application of Recombinant DNA in Medicine
1. With help of rDNA, many approaches towards medicine has been evolved, with help of amplification and
vectors, large number of human hormones/ proteins can be engineered in pharmaceutical industry. Like
human Growth hormone, Insulin, Blood clotting factors, Monoclonal Ab, Interferon and vaccine in large
scale.
● Insulin:
Is a hormone responsible for glucose level maintenance, defect in production of quality of insulin leads to
diabetic condition.
Before existence of rDNA, animals were slaughtered and insulin was isolated.
Now, with help of rDNA it is possible to isolate human insulin gene and put in E.coli→cultured→
formation of desired quantity of insulin which is commercialised.
● Human Growth Hormone:
Is a natural hormone responsible for, growth,differentiation, regeneration in human body, in many patients
dwarfism occurs due to insufficient HGH. Which can be treated by injecting recombinant HGH. In this,
human growth gene is used and transferred to plasmid.

10. ● Interferon:
Is a protein responsible for interfering in virus protein synthesis, when virus is intruded, host body give
response by releasing Interferon. This characterisation is targeted and is used to treat cancers, other virus
caused infection.
Curing: hairy cell leukemia, Alpha component in rDNA cures lymphoma & myelogenous leukemia.
● Hemophilia A/B:
Due to deficiency of blood clotting factor 8 & 9, caused by mutation or inheritance of defect in X-
chromosome, hemophilia can be observed. Which can be overcome by recombinant 8 & 9 clotting factors.
● Antibiotics:
1st example of antibiotic is penicillin which was developed by Alexander Fleming in 1928. It was
artificially prepared by recombinant DNA technology by denaturing harmful living pathogens.
● Antibodies are specific proteins produced by the immune system in response to presence of specific
antigen.
1. Monoclonal antibodies.
Used in diagnostic agent of infection and pregnancy test
Vaccine effectiveness developed for human animal and plants.
Monoclonal antibodies are produced from single clone of antigen and which is, why monospecific in
nature.
Is developed with hybridoma Technology.

11. ● Vaccine:
Vaccine are live attenuate (but suppressed or dead), which triggers immunogenic response towards
inserted antigen forming antibodies (defence army) which helps to fight with real antigen when exposed.
With help of rDNA, cloning of protective gene
can be used to form antibodies against antigens.
● Examples: HIV, Hepatitis B, Herpes,
Influenza, Mouth and foot disease.

13. 2. Diagnosis:
Molecular diagnosis of infectious disease was mainly depended on isolation and identification which was a
lengthy and slow process .Meanwhile development of diagnostic kits to identify infectious pathogenic organisms
by knowing the specific DNA sequence of organism led to accurate and fast results.
● Diagnostic Kits of AIDS, cancer, foot and mouth disease tuberculosis etc is now available.
Majorly it is based on Elisa, PCR and RIA essay principles.
For example in the diagnosis of HIV: "first antibody test will proceed by using recombinant HIV protein to
measure antibodies in body that proliferate when there is HIV infection. Then, the DNA test uses reverse
transcription polymerase chain reaction (RT-PCR) to detect presence of HIV genetic material. This is used for
analyzing the genome sequence." Constructing probes "that are short sequence nucleotides" with labelled marker
are important strategy in diagnosis of diseases while other is MAB production.
3. Gene Therapy:
Many of times Genes are mutated or defected rising diseased condition, to overcome Gene can be manipulated
or healthy gene is cultured in vector and then injected in patient.
Which can be done in- vivo/ ex-vivo

15. rDNA in Forensic industry:
1. DNA Fingerprinting:
-The DNA from a biological sample left at the scene of a crime can be analyzed and compared with
the DNA of likely suspects.
-The chemical structure of every organism’ DNA is the same. The only difference between
organisms is the order of the basic pairs.
● Establishment of Child percentage
● Evaluation of trace left by criminals
● Deduction of racial groups

16. Thank you for patience listening.