Construction and expression of anti- HIV-1 ScFv and diphtheria toxin fusion gene in E. coli

Objective To construct the expression vector of anti-HIV-1 ScFv and diphtheria toxin fusion gene and induce it expresses fusion protein in E coli. Methods DAB389 gene fragment obtained by PCR and clone into prokaryotic expression vector pET-28a, and then transform into E coli. BL21 (DE3) for expression under induction of IPTG. The expression product was identified by SDS-PAGE and Western blotting. Results Enzyme digestion analysis and PCR identification shown that the recombinant plasmid was constructed correctly. SDS-PAGE and Western blot analysis indicated that the Mr 75200 protein in a form of inclusion body was expressed, which was the same as the fusion gene DT-hS120. The targeted expression protein was about 21% of total protein. Conclusion The expression vector of DT-hS120 fusion protein is constructed successfully, and procured high performance expression.