Overexpression in Escherichia coli, Purification and Characterization of Thermoanaerobacter tengcongensis Elongation Factor G with Multiple Rare Codons

Title: Overexpression in Escherichia coli, Purification and Characterization of Thermoanaerobacter tengcongensis Elongation Factor G with Multiple Rare Codons

Volume: 14 Issue: 8

Author(s): Liqiang Zhang, Peng Xue and Hongjie Zhang

Affiliation:

Keywords: Elongation factor G, RIG plasmid, gene expression, purification and characterization, Thermoanaerobacter tengcongensis

Abstract: The fusA gene encoding a thermophilic protein EF-G with multiple rare condons was cloned from Thermoanaerobacter tengcongensis (TteEF-G) and overexpressed in Escherichia coli by cotransfering a RIG plasmid to overcome the potential codon-bias problem originated from Arg, Ile and Gly. The recombinant protein was identified by MALDITOF- MS for molecular mass with approximation of 76 kDa and by trypsin digestion coupled LC-MS/MS for peptide sequence coverage of 61.3%. The in vivo complementary assay indicates that TteEF-G could significantly rescue the E. coli LJ14 (frrts) at the non-permission temperature of 42°C in the bi-transformant of TteRRF and TteEF-G. This study indicated that coexpression of rare codons cognate tRNA is a useful method for protein overexpression in E. coli.

Cite this article as:

Liqiang Zhang , Peng Xue and Hongjie Zhang , Overexpression in Escherichia coli, Purification and Characterization of Thermoanaerobacter tengcongensis Elongation Factor G with Multiple Rare Codons, Protein & Peptide Letters 2007; 14 (8) . https://dx.doi.org/10.2174/092986607781483732

DOI https://dx.doi.org/10.2174/092986607781483732	Print ISSN 0929-8665
Publisher Name Bentham Science Publisher	Online ISSN 1875-5305