[Cloning, expression of Schistosoma japonicum elastase gene and its stage-specific transcription].
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi
; 25(5): 359-63, 2007 Oct.
Article
en Zh
| MEDLINE
| ID: mdl-18441984
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.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.
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Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Schistosoma japonicum
/
Elastasa Pancreática
/
Proteínas Protozoarias
/
Perfilación de la Expresión Génica
Límite:
Animals
Idioma:
Zh
Revista:
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi
Asunto de la revista:
PARASITOLOGIA
Año:
2007
Tipo del documento:
Article
País de afiliación:
China