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


CONCLUSIONS
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.
National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention; Laboratory of Parasite and Vector Biology, Ministry of Health, China.
Ying
Y
Wang
Jing
J
Zhang
Zhong-Ying
ZY
Yuan
He-Jun
HJ
Zhou
Yu-Juan
YJ
Shen
Yu-Xin
YX
Xu
Yan-Juan
YJ
Wang
Wei-Ping
WP
Wu
Jian-Ping
JP
Cao
D000818:Animals; D003001:Cloning, Molecular; D004443:Echinococcosis / Q000175:diagnosis* / Q000276:immunology / Q000469:parasitology; D048209:Echinococcus granulosus / Q000201:enzymology* / Q000235:genetics / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D015801:Helminth Proteins / Q000235:genetics* / Q000276:immunology; D010751:Phosphopyruvate Hydratase / Q000235:genetics* / Q000276:immunology; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology
D015801:Helminth Proteins; D011994:Recombinant Proteins; D010751:Phosphopyruvate Hydratase