Synonymous rare arginine codons and tRNA abundance affect protein production and quality of TEV protease variant.

ABSTRACT: It has been identified that a TEV protease (TEVp) variant, TEVp(5M), displays improved solubility. Here, we constructed fifteen TEVp(5M) variants with one or more of six rare arginine codons in the coding sequence replaced with abundant E. coli arginine codons. These codon variants expressed in either E. coli BL21 (DE3) or Rossetta (DE3) showed different solubility and activity. Supply of rare tRNAs increased the tendency of certain codon variants to form insoluble aggregates at early induction stage, as determined by the fused S-tag. About 32% increase in soluble protein production of M5 variant with four synonymously mutated arginine codons was identified in Rossetta (DE3) cells using GFP fusion reporter, comparable to that of TEVp(5M). After purification, two other codon variants from b

SUBMITTER: Fang J 

PROVIDER: S-EPMC4245098 | biostudies-literature | 2014

REPOSITORIES: biostudies-literature