Construction and Prokaryotic Expression of Anti-HIV-1 Diphtheria Toxin Immunotoxin

Objective:To construct the expression vector for DTA-hS120 fusion protein and induce the fusion protein express in E.coli.Methods:Amplify the DTA gene fragment only contain the active site of diphtheria toxin by PCR and clone into prokaryotic expression vector pET-28a,then transform into E.coli BL21(DE3) for expression under induction of IPTG.Identify the expression product by SDS-PAGE and Western blot.Results:The recombinant plasmid was identified by incide and gene sequencing.SDS-PAGE and Western blot analysis showed that a protein about 54 kD in a form of inclusion body was expressed,which contained about 23% of total somatic protein.The molecular weight of this protein was the same as the fusion protein DTA-hS120.Conclusion:The expression vector of DTA-hS120 fusion protein was constructed,and procure high performance expression.