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
Open Access
- 2 September 1999
- journal article
- Published by Wiley in FEBS Letters
- Vol. 457 (3) , 393-396
- https://doi.org/10.1016/s0014-5793(99)01081-9
Abstract
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 tRNAAGA, tRNAAGG 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.Keywords
This publication has 17 references indexed in Scilit:
- Cloning of the Gene for Inorganic Pyrophosphatase from a Thermoacidophilic Archaeon,Sulfolobussp. Strain 7, and Overproduction of the Enzyme by Coexpression of tRNA for Arginine Rare CodonBioscience, Biotechnology, and Biochemistry, 1998
- Molecular chaperones, folding catalysts, and the recovery of active recombinant proteins fromE. coliApplied Biochemistry and Biotechnology, 1997
- High-level Misincorporation of Lysine for Arginine at AGA Codons in a Fusion Protein Expressed inEscherichia coliJournal of Molecular Biology, 1996
- A modified uridine in the first position of the anticodon of a minor species of arginine tRNA, the argU gene product, from Escherichia coliEuropean Journal of Biochemistry, 1993
- Genomic organization and physical mapping of the transfer RNA genes in Escherichia coli K12Journal of Molecular Biology, 1990
- Frameshift suppression at tandem AGA and AGG codons by cloned tRNA genes: assigning a codon toargUtRNA and T4 tRNAArgNucleic Acids Research, 1990
- High-level expression of recombinant genes in Escherichia coli is dependent on the availability of the dnaY gene productGene, 1989
- Mistranslation in IGF-1 during over-expression of the protein in Escherichia coli using a synthetic gene containing low frequency codonsBiochemical and Biophysical Research Communications, 1988
- The E. coli dnaY gene encodes an arginine transfer RNACell, 1986
- Correlation between the abundance of Escherichia coli transfer RNAs and the occurrence of the respective codons in its protein genesJournal of Molecular Biology, 1981