High level expression of Thermococcus litoralis 4-alpha-glucanotransferase in a soluble form in Escherichia coli with a novel expression system involving minor arginine tRNAs and GroELS.
The Thermococcus litoralis 4-alpha-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.
Imamura
H
H
Department of Biotechnology, The University of Tokyo, Yayoi, Bunkyo-ku, Tokyo, Japan.
Jeon
B
B
Wakagi
T
T
Matsuzawa
H
H
D018834:Chaperonin 60 / Q000235:genetics* / Q000378:metabolism; D003062:Codon; D004926:Escherichia coli / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial; D006004:Glycogen Debranching Enzyme System / Q000235:genetics* / Q000378:metabolism; D015202:Protein Engineering / Q000379:methods; D012347:RNA, Transfer, Arg / Q000235:genetics*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D018741:Replication Origin; D012995:Solubility; D019712:Thermococcus / Q000201:enzymology* / Q000235:genetics
D018834:Chaperonin 60; D003062:Codon; D006004:Glycogen Debranching Enzyme System; D012347:RNA, Transfer, Arg; D011994:Recombinant Proteins; C020569:4 alpha-glucanotransferase