What steps are involved in the synthesis of genetically engineered insulin?
The steps involved in the production of genetically engineered insulin are given below: RNA is extracted from the beta-cells of Islet of Langerhans in the pancreas to synthesize insulin. Single-stranded DNA complementary to mRNA is synthesized with the help of the enzyme reverse transcriptase. Then the second chain of DNA complementary to the first DNA sequence is synthesized with the help of enzyme DNA polymerase. Both the replicated DNA strands are joined with the plasmid using a terminal transferase enzyme. The enzyme named ligase is used to anneal both ends of the DNA. In this way, the ends of the plasmid and inserted DNA are sealed to form a new circular plasmid. This is a recombinant DNA molecule. In the next step, this recombinant DNA is inoculated into new E.coli bacterial cells, digested with restriction enzymes, and inserted into the bacterial gene. After adequate gene expression, the germ cells from both cultures are lysed with the appropriate chemical. The insulin portion is then separated from the enzyme using cyanogen bromide
