Cloning, expression and purification of new encoding gene of GST from clonorchis sinensis

  Objective   To amplify a new encoding gene of GST from adult C.sinensis(CsGST gene), determine whetherithad intron or not, express it in Escherichia coli(E.coli)and purify the recombinant protein.

  Methods   The gene encoding Cs GST was obtained from the cDNA(plasmid)library and genomic DNA of adult C.sinensis by PCR and sequenced.The coding gene was cloned into the prokaryotic expression vector p ET-30a(+)and expressed in E.coli BL21/DE3.The recombinant protein was purified according to the protocol of Ni-NTA agarose(QIA GEN, Germany).It was analyzed by SDS-PAGE and TLC.

  Results   The full length of the gene encoding CsGST was 639 bp.Ithad no intron.The recombinant plasmid p ET-30a(+)-CsGST was efficiently expressed in E.coli, its molecular weight was about 30 kDa.The recombinant protein occupied 33% of total bacterial protein.The purity of the purified recombinant protein was 97%.

  Conclusion   The CsGST gene of C.sinensis had no intron.Its efficient expression was achieved in E.coli.The recombinant protein was purified.These can lay a base for its function research.