Recombinant DNA Technology and its Applications.. 1)DNA 2)r-DNA Technology 3)HISTORY 4) ENZYMES IN r-DNA TECHNOLOGY 5)APPLICATION 6)INSULIN 7)HEPATITIES 8) ADVANTAGES & DISADVANTAGES... PRESENTED BY : MR. ARINDAM CHAKRABORTY, M.PHARM IN PHARMACEUTICAL BIOTECHNOLOGY, CALCUTTA INSTITUTE OF PHARMACEUTICAL TECHNOLOGY & AHS ULUBERIA, HOWRAH , WEST BENGAL , PIN 711316

1. APPLICATION OF r-DNA
TECHNOLOGY IN
DEVELOPMENT OF NEW
DRUG
ARINDAM CHAKRABORTY
M. PHARM, 1ST SEMESTER
DEPARTMENT OF PHARMACEUTICAL
BIOTECHNOLOGY
ROLL NO : 15920919003
CALCUTTA INSTITUTE OF PHARMACEUTICAL
TECHNOLOGY & A. H. S

2. CONTENTS
 DNA
 r-DNA TECHNOLOGY
 HISTORY
 ENZYMES IN r-DNA TECHNOLOGY
 APPLICATION
 APPLICATION IN NEW DRUG
 INSULIN
 SYNTHESIS OF INSULIN
 HEPATITIS –B
 SYNTHESIS OF HEPATITIS –B
 ADVANTAGE AND DISADVANTAGE
 REFERENCE

3. D.N.A
Deoxyribonucleic acid, more commonly known as DNA, is a
complex molecule that contains all of the information necessary
to build and maintain an organism.
 All DNA base made up of a base consisting a sugar
phosphate and nitrogen base
 It consists of double helix
 Contains Four nitrogen bases:
1. Adenine
2. Guanine
3. Thymine
4. Cytosine

4. r-DNA TECHNOLOGY
Recombinant DNA or r-DNA are the artificial DNA created by
combining two or more sequences of DNA. Production of a unique
DNA molecule by joining together or more DNA fragments not
normally associated with each other, which can replicate in the
living cell.
 Recombinant DNA is also called chimeric DNA
 TWO ENZYMES MAINLY TAKES PART ON IT:
1. Restrictions enzyme
2. DNA ligase
 METHODS:
1. Transformation
2. Conjugation
3. Transduction

5. HISTORY
 It was developed by two scientists named Stanley Cohen
and Herbert Boyar in 1972.

6. ENZYMES IN r-DNA
TECHNOLOGY
1. Restriction endonucleas: Cleaves DNA in specific
fragments
2. DNA ligase: Binds two DNA molecules or fragments
3. DNA polymerase: Fills Single strand gaps
4. Reverse Transcriptase: Makes DNA to RNA
5. Polyneucelotide Kinase: Add a phosphate in 5' OH
end.

7. APPLICATION
 DNA technology is also used to detect the presence of HIV in a
person.
 Application of recombinant DNA technology in Agriculture – For
example, manufacture of Bt-cotton to protect the plant against
ball worms.
 Application of medicines – Insulin production by DNA
recombinant technology is a classic example.
 Gene Therapy – It is used as an attempt to correct the gene
defects which give rise to heredity diseases.
 Clinical diagnosis – ELISA is an example where the application
of recombinant

8. APPLICATION IN NEW DRUG
1. Insulin
2. Erythropoietin
3. Hepatitis
4. Interferon

9. INSULIN
 Insulin is a hormone that regulates the amount of glucose
(sugar) in the blood and is required for the body to
function normally. Insulin is produced by cells in the
pancreas, called the islets of Langerhans. These cells
continuously release a small amount of insulin into the
body.
 There are three steps in Insulin Biosynthesis:
 PRE PRO INSULIN(Contains 110 Amino acid)
 PRO INSULIN (Contains 86 Amino acid)
 INSULIN (Contains 51Amino acid)

10. SYNTHESIS OF INSULIN
ISOLATION OF cDNA FROM PANCREAS
CUTTING OF DNA BY RESTRICTION ENZYME
JOINING WITH VECTOR
PLACED INTO THE HOST (E.coli)
DNA LIGASE AND CnBr ADDED
EXTRACTION OF NEW SEQUENCE OF DNA

11. SYNTHESIS OF INSULIN

12. HEPATITIS-B
 Hepatitis B is a serious liver infection caused by the hepatitis B virus
(HBV). For some people, hepatitis B infection becomes chronic,
meaning it lasts more than six months. Having chronic hepatitis B
increases your risk of developing liver failure, liver cancer or cirrhosis
— a condition that permanently scars of the liver.
 SYMPTOMS:
Hepatitis B signs and symptoms may include:
 Abdominal pain
 Dark urine
 Fever
 Joint pain
 Loss of appetite
 Nausea and vomiting

13. SYNTHESIS OF HEPATITIS-B
ISOLATING CDNA FROM SPECIFIC SITES
CUTTING OF DNA BY RESTRICTION ENZYME
JOINING WITH THE VECTOR .
PLACED INTO HOST CELL.(YEAST CELL)
FERMENTATION TANK
CULTURE – PRODUCTION-PURIFICATION.

14. SYNTHESIS OF R-ERYTHROPOIETIN

15. ADVANTAGES
 No need for natural or organic factors.
 Unlimited utilizations.
 Provides substantial quantity.
 Cheap.
 Resistant to natural inhibitors.

16. DISADVANTAGES
 Commercialized and became big source of income for
businessman.
 Effects natural immune system of the body.
 Can destroy natural ecosystem that relies on organic cycle.
 Prone to cause mutation that could have harmful effects.
 Major international concern : manufacturing of biological
weapons such as botulism & anthrax to target humans with
specific genotype.
 Concerns of creating super – human race

17. REFERENCE
 Dr. Dubey R.C. ; “A Text Book of Biotechnology” ;370-373.
 Kammermayor K. ;Clark, V. L. ; “Genetic Engineering Fundamentals, An
introduction to Principles & Applications”; Marcel Decker Inc.; 1989; 502.
 Inoue N; Takeuchi M; Ohashi H; Suzuki T ; “The production of
recombinant human erythropoietin” ;Biotechnology Annual review; 1995;
1(1); 297-313
 Satyanaryana U; Chakrapani U ; “Biotechnology”; 75-78, 189-198.
 Dr. Chandel R; Dr. Chatterjee G; “ Postgraduate biochemistry companion”;
Innovative Publication; 24-56.

18. THANK TOU..