Abstract:
Objective To express on temperature-dependent and constructan prokaryotic expression vector carrying human GSTM1TV2 gene and expression with high efficiency in E.coli DH5a.
Methods The GSTM1TV2 cDNA was amplified from human lung total RNAs by RT-PCR and was recombined with prokarytic expression vector pBV220.The recombined plasmid pBV220-GSTM1TV2 were verified with PCR, restriction analysis and sequencing determination and induced with temperature-depenent in 42℃.
Results The expressed non-fusion protein, with molecular weight of about 26 kDa, was about 33% of the total cell protein by SDS-PAGE. Human GSTM1TV2 was recombined correctly with pBV220.Comparing with Genbank, it is correct.
Conclusion The recombined plasmid pBV220-GSTM1TV2 was constructed for research on toxicology and pharmacogenetics.