[Cloning, expression and immunodiagnostic evaluation of enolase from Echinococcus granulosus].

Abstract

OBJECTIVE:

To clone and express EgEno gene of Echinococcus granulosus, and to investigate the immunogenicity and diagnostic value of recombinant EgEno.

METHODS:

Total RNAS of E. granulosus was extracted and reversedly transcripted to cDNA. EgEno gene was amplified from cDNA and inserted into vector pET28a. The recombinant plasmid pET28a-EgEno was transformed into E. coli BL21 (DE3) for expression under the induction of IPTG. The expressed product was identified by SDS-PAGE and Western blotting. The purified recombinant EgEno protein was detected by ELISA with the sera of cystic echinococcosis patients, healthy persons and other patients.

RESULTS:

The EgEno gene was successfully amplified from cDNA of E. granulosus and a fusion protein was expressed in E. coli BL21 (DE3). The molecular weight of the expressed protein was around 50 kDa. The result of Western blotting indicated that the antigenicity of the protein was specific. The sensitivity of diagnosis by ELISA for cystic echinococcosis was 81.25%.

CONCLUSION:

EgEno of E. granulosus is cloned and expressed in E. coli BL21 (DE3) successfully, which might be used as a candidate antigen of immunodiagnosis for cystic echinococcosis.

• Authors: Ying Wang, Jing Zhang, Zhong-Ying Yuan, He-Jun Zhou, Yu-Juan Shen, Yu-Xin Xu, Yan-Juan Wang, Wei-Ping Wu, Jian-Ping Cao
• Journal: Zhongguo xue xi chong bing fang zhi za zhi = Chinese journal of schistosomiasis control (Zhongguo Xue Xi Chong Bing Fang Zhi Za Zhi) Vol. 24 Issue 5 Pg. 549-52, 556 (Oct 2012) ISSN: 1005-6661 [Print] China
• PMID: 23373261 (Publication Type: Evaluation Study, Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Helminth Proteins
• Recombinant Proteins
• Phosphopyruvate Hydratase

Topics

• Animals
• Cloning, Molecular
• Echinococcosis (diagnosis, immunology, parasitology)
• Echinococcus granulosus (enzymology, genetics, immunology)
• Enzyme-Linked Immunosorbent Assay (methods)
• Helminth Proteins (genetics, immunology)
• Phosphopyruvate Hydratase (genetics, immunology)
• Recombinant Proteins (genetics, immunology)