Difficulty: Medium
Correct Answer: Using chemically synthesized DNA sequences for the A and B chains separately (expressed as fusion proteins and then recombined)
Explanation:
Introduction / Context:
Recombinant human insulin was produced using early bacterial expression strategies that overcame challenges of peptide stability and processing. Understanding the chain-by-chain method explains how bioidentical insulin was obtained without relying on animal pancreases.
Given Data / Assumptions:
Concept / Approach:
One classic strategy used two synthetic genes (for A and B chains). Each chain was expressed in E. coli as a β-galactosidase fusion, cleaved with cyanogen bromide, purified, and then oxidatively refolded together to form native disulfide bonds. Alternative approved methods later used proinsulin cDNA with enzymatic processing, but the chain-by-chain method is historically significant and matches the option given.
Step-by-Step Solution:
Verification / Alternative check:
Regulatory and historical accounts of Humulin describe chain-by-chain and proinsulin strategies; both yield bioidentical insulin but the option specifying separate chain synthesis matches the classic approach.
Why Other Options Are Wrong:
Common Pitfalls:
Assuming bacteria can process preprohormones like human cells; engineered strategies are required to obtain mature insulin.
Final Answer:
Using chemically synthesized DNA sequences for the A and B chains separately (expressed as fusion proteins and then recombined).
Discussion & Comments