High level expression of Thermococcus litoralis 4‐α‐glucanotransferase in a soluble form in Escherichia coli with a novel expression system involving minor arginine tRNAs and GroELS

FEBS Letters

Wakagi, Takayoshi¹  Imamura, Hiromi¹  Matsuzawa, Hiroshi¹  Jeon, Beong-Sam¹ 

[1] Department of Biotechnology, The University of Tokyo, Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan

关键词: Rare codon;    Transfer RNA;    GroELS;    Recombinant protein;    4-α-Glucanotransferase;    Thermococcus litoralis;    GTase;    4-α-glucanotransferase;    tRNAAGA;    transfer RNA specific for AGA codons;    tRNAAGG;    transfer RNA specific for AGG codons;    IPTG;    isopropyl-β-D-thiogalactopyranoside;   

DOI  :  10.1016/S0014-5793(99)01081-9

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The Thermococcus litoralis 4-α-glucanotransferase (GTase) gene has a high content of AGA and AGG codons for arginine, which are extremely rare in Escherichia coli. Expression of the GTase gene in E. coli resulted in low protein production and the accumulation of inclusion bodies. However, simultaneous expression of GTase with tRNA_AGA, tRNA_AGG and GroELS affected both the production and solubility of GTase, and production of soluble GTase increasing about 5-fold. This new E. coli expression system should be applicable to the expression of not only archaeal but also eukaryotic genes, which usually contain a large number of AGA and AGG codons.

【 授权许可】

Unknown