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| Prokaryotic expression and purification of Hepatitis C virus core protein（HCV-C） and preparation of polyclonal antibody against HCV-C |
| CHEN Hui; GAO Na; FAN Dong-Ying; ZHENG Qun; WANG Juan; AN Jing |
| Department of Microbiology, College of Basic Medicine, Capital Medical University, Beijing 100069, China |
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Abstract To investigate the interaction between hepatitis C virus core protein (HCV-C) and liver cells, purified HCV-C was obtained and polyclonal antibodies against HCV-C were prepared in the present study. The plasmid containing HC-J4-91 whole genome of HCV1b subtype was used as the template and the HCV-C gene was TA-cloned by polymerase chain reaction (PCR). The prokaryotic expression plasmid containing 6×His and HCV-C gene was constructed and named as pQE31-HCV-C. After expression in Escherichia coli and purification, the recombinant HCV-C was used to immunize BALB/c mice to obtain the antiserum. The titer and specificity of the antibody were determined by enzyme-linked immunosorbent assay (ELISA), Western blot and indirect immunofluorescence staining. The results demonstrated that the prokaryotic expression plasmid pQE31-HCV-C expressing a 22 000 fusion protein was successfully constructed. ELISA showed that antiserum against HCV-C from immunized mice had high antibody level, with titer at 1:12 800. Immunofluorescence staining and Western blot indicated that the antibody had good specificity and could recognize nature and unnatural HCV-C. It is concluded that the purified HCV-C protein and anti-HCV-C polyclonal antibodies obtained in this study might be useful for studying the molecular interaction mechanism between HCV-C and liver cells.
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Received: 11 August 2010
Published: 25 September 2010
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Corresponding Authors:
AN Jing
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