title: [Cloning, expression of Schistosoma japonicum elastase gene and its stage-specific transcription].
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 sporocysts, with Mr 31000. 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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Biotechnology School, East China University of Science and Technology, Shanghai 200237, China.
authors.forename: Cheng-yu
authors.initials: CY
authors.lastname: Huang
authors.forename: Yan
authors.initials: Y
authors.lastname: Lu
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
authors.forename: Chuan
authors.initials: C
authors.lastname: Ju
authors.forename: Zheng
authors.initials: Z
authors.lastname: Feng
authors.forename: Zhong
authors.initials: Z
authors.lastname: Yang
authors.forename: Sheng-yue
authors.initials: SY
authors.lastname: Wang
authors.forename: Wei
authors.initials: W
authors.lastname: Hu
mesh_terms: D000818:Animals; D015153:Blotting, Western; D003001:Cloning, Molecular; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D015971:Gene Expression Regulation, Enzymologic; D008297:Male; D010196:Pancreatic Elastase / Q000145:classification / Q000235:genetics* / Q000378:metabolism; D010802:Phylogeny; D015800:Protozoan Proteins / Q000235:genetics* / Q000378:metabolism; D011817:Rabbits; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012549:Schistosoma japonicum / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development; D014158:Transcription, Genetic
chemical_list: D015800:Protozoan Proteins; D010196:Pancreatic Elastase