Why might a eukaryotic protein expressed in E. coli remain non-functional?
Expression of eukaryotic proteins in prokaryotic host Escherichia coli often yields inactive product due to multiple physiological mismatches. Codon bias differs; rare eukaryotic codons cause ribosome stalling, premature termination, and truncation. Prokaryotes lack endoplasmic reticulum machinery for N-glycosylation, disulfide isomerization, and other post-translational modifications essential for stability and activity. Molecular chaperone systems and folding environments also differ, leading to misfolding, aggregation into inclusion bodies, and proteolytic degradation. Combined effects expl
Ref: NCERT Biology Class XII Principles on Klenow fill-in labeling, Lehninger Chapter 9 DNA cloning techniques, and Molecular Cloning by Sambrook Chapter 10 documenting end-labeling of cohesive termini.