Expression of HIV-1 recombinant protease and its antibody preparation

SHI Jijing; LIU Chaoqi; LI Bin; WANG Jinjiang; CHEN Jing

Abstract：To prepare HIV- 1 protease protein (PR) and its antibody for studying the biological activity of H IV- 1 PR. The full length gene fragment of HIV-1 PR was amplified by PCR, and then cloned into pET28a (+) prokaryotic expressing vector; the recombinant protein PR was induced by IPTG in E. coli BL21 and the expressed protein was detected by Western blot assay. Subsequently, the purified protein was used to immune rabbits for preparing polyclonal antibody, and then the specificity and titer of the antibody were detected respectively by ELISA and immunohistochemistry. We constructed the plasmid pET28a (+) -PR successfully and expressed HIV-1 PR protein in E. coliBL21 efficiently; the recombinant protein was characterized with His antibody by Western blot analyzing, while the rabbit immunized with the purified protein produced high titer of antibody. The viral protein expressed in eukaryotic cells was specifically combined with the anti-body by immunohistochemistry. All results showed that the expressed recombinant HIV-1 PR possessed high antigenicity, and the prepared polyclonal antibody could be used to detect HIV-1 viral protein that expressed in eukaryotic cells, which provided reliable tools for the fu-tare HIV-1 research.