Expression And Characterization Of Anti-HIV-1 Gp41 DsFv And Recombinant Directing Toxin

The genes of d41 was amplified by PCR-based point mutagenesis strategy,and the gene of staphylococcus aureus extoxin A(SEA′) was obtained by PCR.At last,chimeric gene of recombinant directing toxin sd41 was obtained by inserting SEA′gene into the upstream of gene d41.The gene d41 and sd41 were cloned into plasmid pET-28a(+).Plasmids pET-d41 and pET-sd41 were transformed into E.coli BL21(DE3)and induced by IPTG, the expressed target protein mainly existed in the form of inclusion body. The products were purified by Ni-NTA agarose affinity chromatography.Then the eluted products were renatured successfully.The ELISA was used to detect the active and stability,the actively of dsFv and the recombinant directing toxin sd41 were lower then the 41 and SL41,but the stability raised.The results lay the foundation for the further research on its activity application.