What is the main objective of in vitro mutagenesis?
In vitro mutagenesis is a reverse genetics strategy aimed at generating defined nucleotide changes in cloned genes to investigate relationship between protein primary structure and biological function. By altering specific codons corresponding to active site residues, binding motifs, or structural domains, researchers evaluate effects on catalysis, stability, interaction, or localization. Results link sequence to function, guiding enzyme mechanism analysis and rational protein engineering. Objective is not to measure transcription rate, discover unknown genes randomly, or simply amplify DNA. C
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.
