Cloning and Recombinant Expression modified protective antigen from Bacillus anthracis In E. coil

Anthrax is a common illness between human and animal which the agent of that is Bacillus anthracis. The modified protective antigen cab be used in treatment and vaccination. The aim of this study is the recombinant expression of modified protective antigen in E. coli.
Material and Gene fragments were amplified with PCR from pXOI and fusion gene from SOEing PCR was cloned in a cloning vector and finally sub cloned in pET28a() vector and transferred to E. coli BL21(DE3) competent cells. Recombinant expression of modified PA was induced by IPTG and after protein purification with affinity chromatography, resulted antigen was injected to mice in 4 repeats. Polyclonal antibodies produced in mice serum was accessed.
The modified protective antigen cloned in expression vector pET28a() was confirmed by PCR, enzymatic digestion and sequencing. Recombinant protein was confirmed by SDS-PAGE and western blot. Serum was separated from mice blood and titre of antibody against modified PA was assessed by indirect ELYSA.
resulted protein has high inhibitory property for LF and EF virulence factors. By consideration in PA detection by polyclonal antibody against modified PA from Bacillus anthracis, it can be used in for treatment and prophylaxis against anthrax in individual or combination forms with other PA domains.