Expression, purification and characterization of the mixed total-OMP-CagA from Brucella abortus and Helicobacter pylori as vaccine Candidate

Abstract

Brucella can causes brucellosis in humans and animals, the Most significance of this bacterium is due to its use in biologic wars and bioterrorism. Vaccination strategy has played a high role in decreasing Brucella infections in many countries of the world. So, this study aimed to investigate the expression, purification and characterization of the mixed Total-OMP-CagA from Brucella abortus and H. pylori as vaccine candidate. After bioinformatics designing by suitable softwares, OMPs proteins of B. abortus extracted, and recombinant protein CagA that cloned in Pet28a, previously, transformed to the E. coli BL21 as expression host. Then recombinant protein purified by nickel column. Then, CagA and OMPs proteins confirmed with SDS-PAGE and western blotting. In this study construct of Total OMP-CagA was synthesized. OMPs proteins with sizes 25-27 and 36-38 kDa were extracted. Protein 32 KDa of CagA with concentration 700μg/ml expressed and purified, successfully. In this study, the recombinant protein CagA of H. pylori successfully expressed and total OMPs of B. abortus were successfully extracted and combined together to construct a vaccine candidate. But, complementary studies are required to evaluate the immunological features of mixed OMPs-CagA as novel and efficient vaccine candidate against H. pylori.