Explain the steps involved in production of humulin .

Explanation

In mammals , including humans, insulin is synthesized as a pro - horome (like a proenzyme , the prohomone also needs to be processed before it become a fully nature and functional hormone) which contains an extra stretch called the C peptide. This C peptide is not present in the maturation into insulin. The main challenge for production of insulin using rDNA technique was getting insulin assembled into a mature form . In 1983 , Eli Lilly an American company , first prepared two DNA sequences corresponding to A and 6 chain of human insulin and introduced them in plasmids of Escherichia coil to produce insulin chains . Chains A and B were produced separately, extracted and combined by creating disulfide bonds to form human insulin (humulin).


Molecular structure of insulin was worked out by Sanger. Tsan synthesised human insulim first time. Insulin is not administered orally to the diabetic persons.
Production of Human Insulin. Gene transfer involves essentially the following stages -
1. Isolatin of Donor or DNA segment. A useful DNA segment is isloated from the donor organism.
2. Formation of Recombined DNA (DNA). Both the vector and donor DNA segment are cull in the presence of restriction endonulclease. In the presence of ligase DNA segments of both ard! joined to form INDA.
3. Production of Multiple Copies of rDNA. Next step in the process is production of multiple copies of this recombinant DNA.
4. Introduction of DNA in the recipient organism. This rDNA is inserted info a recipient organism,
5. Screening of trie transformed cells. The recipient (host) cells are screened in the presence of DNA and the product of donor gene. The transformed cells are separated and multiplied. All economical method for its mass production. The various steps and their sequence for the production of human insulin are depicted in Fig.