molecular genetics -human insulin production

Question 1

1: isolating the gene

Answer 1

human insulin gene is isolate from a human pancreas cell. a restriction enzyme cuts the restriction sites of the gene and sticky ends are produced at both ends of the gene

Question 2

2: manipulation of bacterial plasmid

Answer 2

a plasmid is removed from a bacterium, and is cut with the same restriction enzyme. sticky ends produced are complementary to those of the insulin gene

Question 3

3: insertion of insulin gene into the bacterial plasmid

Answer 3

3:DNA liga de seals the sticky ends of the insulin gene with the cut plasmid and a recombinant plasmid is formed

Question 4

4:recombinant plasmid inserted into bacterium

Answer 4

a recombinant plasmid is inserted into Ecoli bacterium using heat shock method, a transgenic bacterium is produced

Question 5

5: expression of target gene

Answer 5

expression of insulin gene in the bacteria results in production of insulin protein. bacteria is grown in fermenter in optimal conditions for mass production. insulin is then extracted and purified