Expression and self-assembly of HIV-1 CAP2NC protein.

Shimeng Bai; Zhiqing Zhang; Jiaming Qiao; Honglin Shen; Fang Huang; Shuangquan Gao; Shaoyong LI; Shaowei LI; Ningshao Xia; Ying GU

Return

Expression and self-assembly of HIV-1 CAP2NC protein.

Author: Shimeng BAI ¹ ; Zhiqing ZHANG ² ; Jiaming QIAO ² ; Honglin SHEN ¹ ; Fang HUANG ¹ ; Shuangquan GAO ² ; Shaoyong LI ² ; Shaowei LI ¹ ; Ningshao XIA ¹ ; Ying GU ¹
Author Information

1. National Institute of Diagnostics and Vaccine Development in Infectious Disease, School of Life Sciences, Xiamen University, Xiamen 361102, Fujian, China.
2. State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Public Health, Xiamen University, Xiamen 361102, Fujian, China.
Publication Type:Journal Article
Keywords: CAP2NC protein; HIV-1; self-assembly property
From: Chinese Journal of Biotechnology 2018;34(4):586-593
CountryChina
Language:Chinese
Abstract: We constructed the CAP2NC prokaryotic expression vector of HIV-1 NL4-3 strain and obtained relatively pure CAP2NC protein by optimizing its purification conditions to explore its in vitro self-assembly conditions. Primers were designed according to the CAP2NC DNA sequence of HIV-1 NL4-3 strain. The target gene was amplified by PCR and cloned into prokaryotic expression vector pTO-T7. Then the recombinant strain was transformed into Escherichia coli BL21 (DE3). IPTG induced protein expression, then the protein was purified by hydrophobic chromatography. SDS-PAGE and Western blotting were performed to analyze the target protein, and the biological activity of the antigen was identified through ELISA. The self-assembly of CAP2NC protein was analyzed by transmission electron microscopy and gel filtration chromatography. The protein had good reaction with the specific antibodies of p24 and formed different structures in various conditions. When 10% yeast RNA was added to the protein complex, the recombinant protein only formed into a tubular structure, which was similar to the self-assembled structure of the HIV-1 virus capsid. The results showed that the HIV-1 CAP2NC protein had in vitro self-assembly activity, and the RNA affected the structure of CAP2NC protein assembly. The protein can be used as a simple and effective molecular model to study its structure, and then it can provide a reference for the study of HIV immature virus particles.