Article

[Human liver glutathione-S-transferase M1 gene cloning and temperature-dependent expression in E. coli].

Huaxi School of Public Health, Sichuan University, Chengdu 610064, China.
Wei sheng yan jiu = Journal of hygiene research 05/2003; 32(3):236-9. pp.236-9
Source: PubMed

ABSTRACT In order to study the temperature-dependent expression of human liver glutathione-S-transferase M1 gene in prokaryotic cells, the recombinant of prokaryotic expression vector pBV220 with human glutathione-S-transferase M1 gene was constructed. The recombined plasmid pBV220-hGSTM1 was verified with PCR, restriction analysis and sequencing. It's expression was induced with temperature-dependent in 42 degrees C and the expressed non-fusion protein in HD5 alpha, with molecular weight of about 28 kDa, was about 28.3% of the total cell protein determined by SDS-PAGE. The sequence of human liver glutathione-S-transferase M1 gene was verified to be correctly recombined with pBV220 compared with the same sequence in Gene Bank, and code 619 C-->A, the amino acid changed from Pro to Thr was observed. The recombined plasmid pBV220-hGSTM1 may be applicable in toxicological and pharmacogenetical studies.

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Keywords

amino acid
 
expressed non-fusion protein
 
Gene Bank
 
human glutathione-S-transferase M1 gene
 
human liver glutathione-S-transferase M1 gene
 
It's expression
 
pharmacogenetical studies
 
prokaryotic cells
 
prokaryotic expression vector pBV220
 
recombined
 
recombined plasmid pBV220-hGSTM1
 
restriction analysis
 
temperature-dependent expression
 
total cell protein