Site-directed mutagenesis of the aspartokinase gene lysC and its characterization in Brevibacterium flavum
Related Experiment Video
Updated: Oct 6, 2026

Mutagenesis and Functional Selection Protocols for Directed Evolution of Proteins in E. coli
Published on: March 16, 2011
1Culture Collection and Research Center, Food Industry Research and and Development Institute, Hsinchu, Taiwan.
Abstract:
Using overlap extension polymerase chain reaction (PCR), five transformants of Escherichia coli containing site-directed mutagenized lysC beta gene were generated and analysed. Exchange of C to A and C to T at nucleotide 1118 of the mutated lysC beta gene causes a substitution of serine301 in the wild-type enzyme for tyrosine301 and phenylalanine301 in the mutant enzymes, respectively. Enzyme assays showed that Brevibacterium flavum cells harbouring pSUMN18 with mutated lysC beta genes exhibited 16-20 fold lower specific activities of aspartokinase as compared to that of host containing wild-type lysC gene. The mutation introduced into lysC beta of B. flavum CCRC 18271 resulted in partial feedback-resistant aspartokinase activity.
More Related Videos
Related Concept Videos
In-vitro Mutagenesis (Gene Knockouts)
Conservative Site-specific Recombination and Phase Variation
The recognition sites for Cre recombinase called LoxP...
In vitro Mutagenesis

