Expression of recombinant protein VP1and its application in developing enzyme-linked immunoassay for Echovirus 30

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Author:
ZHU Jian-sheng(Department of Infectious Diseases, Taizhou Hospital,Taizhou 317000, Zhejiang Province, China)
RUAN Xian-li(Department of Infectious Diseases, Taizhou Hospital,Taizhou 317000, Zhejiang Province, China)
WANG Zhi-jun(Department of Infectious Diseases, Taizhou Hospital,Taizhou 317000, Zhejiang Province, China)
ZHOU Lin-fu()
WANG Xin-xin(Department of Infectious Diseases, Taizhou Hospital,Taizhou 317000, Zhejiang Province, China)
WANG Li-zhen(Department of Infectious Diseases, Taizhou Hospital,Taizhou 317000, Zhejiang Province, China)
CHEN Hua-zhong(Department of Infectious Diseases, Taizhou Hospital,Taizhou 317000, Zhejiang Province, China)
WANG Geng-ge(Department of Infectious Diseases, Taizhou Hospital,Taizhou 317000, Zhejiang Province, China)
Journal Title:
Chinese Journal of Clinical Infectious Diseases
Issue:
Volume 05, Issue 02, 2012
DOI:
10.3760/cma.j.issn.1674-2397.2012.02.005
Key Word:
Recombinant proteins;Enzyme-linked immunosorbent assay;Echovirus

Abstract: Objective To express VP1recombinant protein of Echovirus 30 (ECHO30) in E.coli BL21(DE3) and to develop an enzyme-linked immunoassay (EIA) based on the recombinant antigen for detecting antibodies to ECHO30.Methods The target VP1gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR).The PCR products of the gene were inserted into pET44a vector,and then expressed in E.coli BL21( DE3 ).The purified recombinant protein was used for the development of EIA.Results The molecular weight of the recombinant protein was 95 000,and the antigenicity of which was identified by Western blot and EIA.Conclusion The recombinant protein VP1has been successfully expressed and purified,which can be used as diagnostic antigen.

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