Polyclonal antibody preparation and expression in liver tissues of transactivated protein 5 of hepatitis C virus nonstructural 5A / 西安交通大学学报·英文版

ABSTRACT

Objective: To prepare polyclonal antibody of transactivated protein 5 of hepatitis C virus nonstructural 5A (NA5ATP5) and to explore its expression in the liver tissues. Methods: In Escherichia coli BL21, the prokaryotic expression vector pET32a(+)-NS5ATP5 was induced by isopropyl-β-D-thiogalactoside (IPTG), and it was analyzed with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting. And the purified protein was used to immunize the rabbit to prepare polyclonal antibody, with which we studied the function of NS5ATP5 by determining the different liver tissues with the streptavidin-perosidase (SP) immunohistochemistry method. Results: Recombinant NS5ATP5 (molecular weight, 65 kD) and polyclonal antibody were successfully prepared. NS5ATP5 expression in the liver of patients with chronic HCV infection was much higher than that of a normal person, and it was detected mainly in the cytoplasm. Conclusion: The findings of the expression difference between HCV patients and normal people led to a novel diagnostic marker to detect HCV infection.