Abstract: The humanize anti-HIV-1 single chain Fv fragment(ScFv) gp41 gene and super antigen staphylococcal exterotoxin A(SEA) gene were fused via soft linker peptide, and cloned into expression vector pPIC9k. The recombinant plasmid was linearized and transferred into Pichia pastoris strains GS115 by electroporation. High copies of trransformants were obtained with Mut^s and Mut⁺ phenotype identification, PCR amplification and screening of G418. After flask culture and inducing expression by methanol, the targeted protein was well expressed and performed via SDS-PAGE and Western Blot. The highest production was 47.9 mg/L when different parameters were optimized. The primarily-purified protein was analyzed with antibody affinity assay, cellular binding activity and cellular killing activity using the HIV-1 infection target cell models prepared by our laboratory. The results suggest that the constructed anti-HIV-1 gp41 recombinant agent SL41 has a good biological activity and specific CTL cytotoxicity activity. This research will provide a potential values for AIDS treatment and the settled solid foundation for clinical trials.

Key words: HIV-1, Recombinant directing agent, Single chain Fv fragment, Staphylocoecus extoxin A(SEA), Secrete expression, Pichia pastoris