中国生物工程杂志

Human immunodeficiency virus (HIV) is the etiologic agent of AIDS.Immunogenic epitopes of HIV resides on virus-encoded envelope glycoproteins. To prepare HIV-1 gp160 protein and used it for clinical diagnosis, three gene fragments of HIV-1 env containing highly immunogenic epitopes were amplified by PCR from plasmid of HIV-1 HXB2. The resulting PCR products were linked and cloned into a prokaryotic expression vector pET28a(+) and the accuracy of the inserted fragments was identified by sequencing. To express the HIV-1 gp160 fusion epitopes in E. coli cells and identify fusion protein, the induced protein was checked with SDS-PAGE and Western Blot (WB). The identified HIV positive serum samples were tested by Western Blot (WB) to analysis immunogenicity of the purified protein. The length of the chimeric fragment derived from HIV-1 gp160 was 969bp, and encoded 37kDa amino acid residues. Procaryotic expression plasmid was constructed successfully which can highly effectively express gp160 fusion protein. Recombinant fusion protein was expressed in Eserichia coli BL21(DE3) as an insoluble protein. Procaryotic expression plasmid which can highly effectively express gp160 fusion protein was constructed successfully. The founding and subjects were provided for the research of diagnosis kit.