Isolation, cloning and expression of the Brucella melitensis Omp31 gene

Document Type: Short paper

Authors

1 Department of Biotechnology, Razi Vaccine and Serum Research Institute, Mashhad, Iran

2 Department of Biotechnology and Plant Breeding, College of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran

3 Department of Microbiology, School of Science, Alzahra University, Tehran, Iran

4 Department of Brucella Vaccine Research and Production, Razi Vaccine and Serum Research Institute, Karaj, Iran

5 Immunology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran

Abstract

Brucellosis is a zoonotic disease transmitted to humans either from animals or from their products.
Although brucellosis can be found worldwide, the Mediterranean Basin, South and Central America, Eastern
Europe, Asia, Africa, the Caribbean, and the Middle East have currently been listed as high-risk regions. The
genus Brucella is classified in at least nine species. Brucella melitensis is the global pathogenic species of
Brucella. The outer membrane protein 31, (Omp31) from B. melitensis is considered as a protective
immunogen and an important candidate vaccine. Contamination of purified Omp31 protein by biochemical
methods has made some restrictions in practical experiments. In this study, the Omp31coding gene of B.
melitensis Rev 1 strain was inserted in pET32b(+) plasmid with extra His-tag sequence. The integrity of the
constructed plasmid was confirmed using restriction enzyme mapping and sequencing. Omp31 was
expressed after induction with IPTG in Escherichia coli BL21. Recombinant Omp31 (rOmp31) was purified
by chromatography through Ni-agarose. The electrophoresis showed successful purification and
immunoblotting confirmed immunereactivity of rOmp31. Obtained rOmp31 could be used as a research
experimental tool in protection assays to find its potential as a vaccine candidate.

Keywords