[Cloning and expression of the F0-ATP synthase B chain of Clonorchis sinensis and immunogenicity identification of the recombinant protein].
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi
; 25(5): 390-3, 2007 Oct.
Article
em Zh
| MEDLINE
| ID: mdl-18441991
ABSTRACT
OBJECTIVE:
To clone and express the Clonorchis sinensis F0-ATP synthase b chain (CsF0-ATP-synt_B) gene and analyze immunogenicity of the recombinant protein.METHODS:
The coding region F0-ATP synthase b chain gene with the mitochondrial targeting sequence (MTS) removed was amplified with PCR using the cloned plasmid as template, and the product was cloned into the prokaryotic expression vector pET-28a(+), transformed into E. coli BL21 (DE3) and induced with IPTG. The expressed product was purified by Ni-IDA affinity chromatography,and analyzed by SDS-PAGE for its expression and identified by Western blotting for its immunogenicity.RESULTS:
The coding sequence of the F0-ATP synthase b-chain like gene removed off the MTS contains 813 base pairs encoding 271 amino acids with a theoretical molecular weight of 31,171.9. PCR, double enzyme digestion and DNA sequencing confirmed that the recombinant plasmid pET-28a (+)-CsF0-ATP-synt_B was constructed successfully, and the resolvable expression was obtained in E.coli BL21. Highly purified recombinant protein was prepared through affinity chromatography. The recombinant protein could be recognized by the immune serum of the SD rat immunized with the recombinant protein.CONCLUSION:
The CsF0-ATP-synt_B like gene has been efficiently expressed in prokaryotic expression system with immunogenicity.
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Base de dados:
MEDLINE
Assunto principal:
Proteínas de Helminto
/
Clonorchis sinensis
/
ATPases Translocadoras de Prótons
Tipo de estudo:
Diagnostic_studies
Limite:
Animals
Idioma:
Zh
Revista:
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi
Assunto da revista:
PARASITOLOGIA
Ano de publicação:
2007
Tipo de documento:
Article
País de afiliação:
China