Step 1: Identification of Gene.
The gene responsible for insulin production is identified from human DNA. Insulin consists of two polypeptide chains – A and B.
Step 2: Insertion into Plasmid.
The insulin gene is inserted into a plasmid vector (circular DNA from bacteria) using restriction enzymes. This recombinant plasmid is then introduced into E. coli bacteria.
Step 3: Expression in Host.
The bacteria, now genetically engineered, produce insulin polypeptides (A and B chains) as they multiply.
Step 4: Extraction and Purification.
The insulin chains produced by bacteria are extracted, purified, and then chemically combined to form functional human insulin.
Step 5: Use in Medicine.
The recombinant human insulin is marketed as "Humulin" and widely used by diabetic patients.
Answer the following questions:
(a) [(i)] Explain how some strains of Bacillus thuringiensis produce proteins that kill certain insects such as lepidopterans but do not kill the Bacillus.
[(ii)] How is the above mechanism exploited for the production of Bt cotton plant by biotechnologists?
(b) [(i)] Explain how the amplification of gene of interest is done using PCR.
[(ii)] State two applications of the desired amplified fragment of DNA.
The basic scheme of the essential steps involved in the process of recombinant DNA technology is summarised below in the form of a flow diagram. Study the given flow diagram and answer the questions that follow.
(a) What is the technical term used for Step 4 in the above process?
(b) Which of the given two combinations of restriction enzyme should be used in Step 1? Justify your answer.
(i) EcoR I to cut the plasmid and Hind III to cut the alien DNA.
(ii) EcoR I to cut both the plasmid and alien DNA.