›› 2007, Vol. 25 ›› Issue (5): 1-363.

• 论著 •     Next Articles

Cloning, Expression of Schistosoma japonicum Elastase Gene and its Stage-specific Transcription

HUANG Cheng-yu1;LU Yan2;WANG Wei1;JU Chuan2;FENG Zheng2;
YANG Zhong1;WANG Sheng-yue3;HU Wei2   

  1. 1 Biotechnology School，East China University of Science and Technology，Shanghai 200237，China；2 National Institute of Parasitic Diseases，Chinese Center for Disease Control and Prevention，WHO Collab-orating Centre for Malaria，Schistosomiasis and Filariasis，Shanghai 200025，China；3 Chinese National Human Genome Center at Shanghai，Shanghai 200231
  • Received:1900-01-01 Revised:1900-01-01 Online:2007-10-30 Published:2007-10-30
  • Contact: HU Wei

PDF (PC)

Abstract:

Objective To clone， express and purify Schistosoma japonicum elastase-2b gene（SjCE-2b）， and analyze its stage-specific transcription. and expression. Methods The coding sequence of the Sj gene was predicted, and a phylogenetic tree of Sj elastase was drawn. RT-PCR and Western blot were used to investigate the differential transcription and expression of SjCE-2b gene during the developmental stages. The SjCE-2b gene obtained by RT-PCR was subcloned into pET28b， and expressed in E.coli （rSjCE-2b）. The expressed protein was purified with His·Tag affinity chromatography. Western blotting was used to investigate the immunogenicity. Results RT-PCR showed specific bands in sporocysts， eggs and adult worms， while Western blot showed that the recombinant protein（rSjCE-2b） existed only in cercariae and sporosysts， with Mr 31 000. The expression vector of SjCE-2b/pET28b was constructed and expressed in E.coli. The recombinant protein rSjCE-2b was specifically recognized by the S. japonicum-infected rabbit serum. Conclusion The transcript of S.japonicum elastase-2b gene was found in sporocysts， eggs and adult worms， and this gene might be a potential candidate for vaccine, for drug and diagnosis target.

Key words: Schistosoma japonicum, Elastase gene, Prokaryotic expression