Some of the steps involved in production of humulin are given below. Choose the correct sequence
(i) Synthesis of insulin gene artificially
(ii) Culturing recombinant E. coli in bioreactors
(iii) Purification of humulin
(iv) Insertion and human insulin gene into plasmid
(v) Introduction of recombinant plasmid into E. coli
(vi) Extraction of recombinant gene product from E. coli.

Options

A. ii, i, iv, iii, v, vi
B. i, iii, v, vi, ii, iv
C. I,iv,v,ii,vi,iii
D. iii,v,ii,I,vi,iv

Answer

To determine the correct sequence of steps involved in the production of Humulin (human insulin), we can break down the process as follows:

1. **Synthesis of insulin gene artificially (Step i)**: The first step in producing Humulin is to synthesize the gene that codes for human insulin. This is essential as we need the specific genetic information to produce the insulin protein.

2. **Insertion of human insulin gene into plasmid (Step iv)**: Once the insulin gene is synthesized, the next step is to insert this gene into a plasmid. Plasmids are small circular DNA molecules that can replicate independently and are commonly used in genetic engineering.

3. **Introduction of recombinant plasmid into E. coli (Step v)**: After the insulin gene is inserted into the plasmid, the recombinant plasmid is introduced into E. coli bacteria. This step is crucial because E. coli will serve as the host organism that will produce the insulin.

4. **Culturing recombinant E. coli in bioreactors (Step ii)**: The next step involves culturing the E. coli in bioreactors. This allows the bacteria to multiply and express the insulin gene, leading to the production of insulin.

5. **Extraction of recombinant gene product from E. coli (Step vi)**: After culturing, the insulin protein needs to be extracted from the E. coli cells. This step involves breaking open the cells and isolating the insulin.

6. **Purification of humulin (Step iii)**: Finally, the extracted insulin needs to be purified to ensure it is suitable for medical use. This involves removing any impurities and ensuring the insulin is in its active form.

Putting these steps together, the correct sequence is:
1. Synthesis of insulin gene artificially (i)
2. Insertion of human insulin gene into plasmid (iv)
3. Introduction of recombinant plasmid into E. coli (v)
4. Culturing recombinant E. coli in bioreactors (ii)
5. Extraction of recombinant gene product from E. coli (vi)
6. Purification of humulin (iii)

Thus, the correct order is: **i, iv, v, ii, vi, iii**.