[Human liver glutathione-S-transferase M1 gene cloning and temperature-dependent expression in E. coli].
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.
Chen
Pingping
P
Huaxi School of Public Health, Sichuan University, Chengdu 610064, China.
Wu
Yiming
Y
Zhang
Chaowu
C
Wu
Yongjun
Y
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005982:Glutathione Transferase / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D006801:Humans; D008099:Liver / Q000201:enzymology*; D010957:Plasmids / Q000235:genetics; D011995:Recombination, Genetic; D013696:Temperature
D005982:Glutathione Transferase