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Virologica Sinica, 16 (3) : 220-223, 2001
Research Article
Expression of Soluble Human SeFv Against Envelope Protein E2 of
Hepatitis C Virus Antigen in E.coli
Gene Theropy Research Center,Institute of infectious,the 302 hospital of PLA,Beijing 100039,China
To construct expressive vector for human ScFv against E2 protein of hepatitis C virus (HCV-E2-ScFv),and express soluble HCV-E2-ScFv in E.coli JM109,using phage display technique,the recombinant phages were panned by recombinant E2 antigen which was coated in a microtiter plate,after five rounds of biopanning,46 clones were identified specific to E2 antigen.750 bp fragment could be released from the plasmid of positive phage colones,and the sequenle analysis indicated that we have got the ScFv BNA fragment.And then DNA fragment was inserted into expressive vector pCANTAB5E and E.coli host JM109 was transformed and induced by IPTG.The specificity of ScFv in the culture medium was evaluated by enzyme-linked immunosorbent assay(ELISA).The molecular weight of expressed HCV-E2-ScFv is 28kD by SDS-polyacrymide gel electrophoresis(PAGE).The expressed HCV-E2-ScFv will be useful in the immunohistochemical study of liver tissue and gene therapy against HCV infectiion.
Key Words: HCV;  E2;  Single chain Fv antibody;  Expression
  Published online: 5 Sep 2001
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