원문정보
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영어
The Escherichia coli has been the most popular host for recombinant protein expressiondue to its rapid growth and well-characterized genetics. It is, however, not unusual that heterologous proteins overexpressed in E. coli often lead to inclusion bodies1). Since the polypeptides being synthesized from the translating ribosomes are exposed to the cytosol vectorially and
co-translationally assisted by the molecular chaperones, the kinetics of protein folding is closely related to the translation initiation and elongation rate2). Thus, to regulate the rate of protein synthesis here we constructed a mutant library with the various translation rates in E. coli by the direct switching of mutant genes with the original target gene in the chromosome. By combination with error-prone PCR, the functions of initiation factor 2 and elongation factor G
could be easily modulated. The mutant library allows the various combinations of translation rates to provide more folding environments. 1) Baneyx, F., and Mujacic, M., (2004) Nat. Biotechnol. 22:1399-1408 2)Frydman, J., (2001) Ann. Rev. Biochem, 70:603-647