Explain how insulin can be produced using recombinant DNA technology.

1.	Explain how insulin can be produced using recombinant DNA technology.
Answer» Insulin Production r-DNA technology 1. A DNA fragment encoding each insulin chain was made by annealing two complementary oligonucleotides that had been chemically synthesised. 2. Each fragments was ligated into a bacterial expression vector such that during translation the insulin chain would be fused to the carboxy terminus of the betagalactosidase enzyme. 3. The expression vectors, were transformed into E.coli and the beta-insulin fusion proteins accumulated inside the bacterial cells. 4. The cells were harvested and each beta-gal-insulin fusion protein was purified. 5. The insulin coding DNA was synthesised so that it started with a methionine codon. This provides a way to cleave off the beta-gal part from the insulin polypeptide. 6. Treatment of the fusion protein with cyanogens bromide (CNBr) cleave the peptides bonds after the methionine. Thus, in this way recombinant insulin is produced in E.coil.

Answer»

Insulin Production r-DNA technology

1. A DNA fragment encoding each insulin chain was made by annealing two complementary oligonucleotides that had been chemically synthesised.

2. Each fragments was ligated into a bacterial expression vector such that during translation the insulin chain would be fused to the carboxy terminus of the betagalactosidase enzyme.

3. The expression vectors, were transformed into E.coli and the beta-insulin fusion proteins accumulated inside the bacterial cells.

4. The cells were harvested and each beta-gal-insulin fusion protein was purified.

5. The insulin coding DNA was synthesised so that it started with a methionine codon. This provides a way to cleave off the beta-gal part from the insulin polypeptide.

6. Treatment of the fusion protein with cyanogens bromide (CNBr) cleave the peptides bonds after the methionine. Thus, in this way recombinant insulin is produced in E.coil.

Explain how insulin can be produced using recombinant DNA technology.

Discussion

No Comment Found

Related InterviewSolutions

Reply to Comment