Construction, Expression and Activities of a Novel Anti-HIV-1 Protein GRFT

A DNA sequence encoding GRFT was synthesized according to the deduced amino acid sequence of GRFT, using an E. colt codon preference table, and cloned into the prokaryotic expression vector pET28a (+). Then the recombinant plasmid was transfered into E. coli BL21 (DE3). The target protein was obtained after induced by isopropyl-beta-D-thiogalactopyranoside (WIG). The results of SDS-PAGE and Western Blot analyses show that the protein is well expressed and had antigenic activities. When different parameters were optimized, the highest production could account for 55. 84% of total bacterial protein. Gel scan analysis showed that the purity was 94. 06% after the target protein refolding and purification with Ni2+-NTA affinity chromatography. The antigenic and cellular biding activities of target protein were identified by Dot-ELISA and TEA methods based on the HIV-1 infection target cell models prepared by our laboratory. The results suggest that the recombinant protein had special recognition and binding activity with target cells. This research will settle solid foundation for further investigate of anti-HIV-1 genetic engineering agents and targeted therapy.