[Expression and activity analysis of the human glutathione-S-transferase in Lactococcus lactis]

Wei Sheng Wu Xue Bao. 2000 Apr;40(2):132-8.
[Article in Chinese]

Abstract

The glutathione-S-transferase A1 cDNA was amplified from human liver total RNAs by RT-PCR and was cloned into a Escherichia coli expression vector pET23b, then the recombinant plasmid pET23bhgst was introduced into E. coli BL21 (DE3) and induced by IPTG, the high-level expression of hGSTA1 appeared in the E. coli cells. The cDNA encoding hGSTA1 was subcloned into pMG36e, a lactococcal expression vector, and introduced into Lactococcus lactis MG1363 by electroporation. In the positive transformants, the hGSTA1 was expressed as a fusion protein which was verified by SDS-PAGE and Western blot. The hGSTA1 from both E. coli and L. lactis was purified by affinity chromatography on glutathione-agarose and all showed enzymatic activity. The potential application of the recombinant Lactococcus lactis in functional food was also discussed.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blotting, Western
  • Cloning, Molecular
  • Electrophoresis, Polyacrylamide Gel
  • Escherichia coli / genetics
  • Gene Expression
  • Glutathione Transferase / biosynthesis*
  • Glutathione Transferase / genetics
  • Glutathione Transferase / metabolism
  • Humans
  • Lactococcus lactis / genetics*
  • Liver / enzymology
  • Plasmids
  • Reverse Transcriptase Polymerase Chain Reaction

Substances

  • Glutathione Transferase