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

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, simultaneo...

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Veröffentlicht in:FEBS letters 1999-09, Vol.457 (3), p.393-396
Hauptverfasser: Imamura, Hiromi, Jeon, Beong-Sam, Wakagi, Takayoshi, Matsuzawa, Hiroshi
Format: Artikel
Sprache:eng
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Zusammenfassung: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.
ISSN:0014-5793
1873-3468
DOI:10.1016/S0014-5793(99)01081-9