Cloning,expression and detection of Hepatitis C virus envelope protein E2
HE Su-mei,ZHOU Jian-yuan,XIAOJian-hua,et al.(1 University of South China,Hengyang,421001,China;2 JYNDA Phamaceutical Co.Ltd,Yueyang,414000,China)
To study the antigenicity and the potential protection of recombinant HCV E2 protein and obtain large amount oftarget product in prokaryotic expression systerms.The HCV E2 gene was amplified by RT-PCR and the product was diges-ted by EcoR I and Sall I.The fragment was inserted into an Escherichia coli expression vector PET41a,and then used totransform E.coli BL21(DE3).The target protein was expressed in BL21(DE3) and induced by IPTG.The expressed pro-tein was indentified by SDS-PAGE and then purified by immobilized metal ion affinity chromatography(IMAC),and thebiological activity of product was detected by ELLSA.The results showed that the prokaryotic expression plasmid of HCVE2 is constructed successfully.The expression product is present in the from of inclusion bodies.The expression productsshow the good reactivity to the positive HCV serum.This result indicate that recombinant protein has high antigenicity,itmay be a useful agent for detecting the anti-HCV antibody.
【CateGory Index】： R373.21