[Cloning, expression of Schistosoma japonicum elastase gene and its stage-specific transcription].
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.
Huang
Cheng-yu
CY
Biotechnology School, East China University of Science and Technology, Shanghai 200237, China.
Lu
Yan
Y
Wang
Wei
W
Ju
Chuan
C
Feng
Zheng
Z
Yang
Zhong
Z
Wang
Sheng-yue
SY
Hu
Wei
W
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
D015800:Protozoan Proteins; D010196:Pancreatic Elastase