cloning, expression and molecular analysis of iranian brucella melitensis omp25 gene for designing a subunit vaccine

brucellosis is a well-known domestic animal infectious disease, which is caused by brucella bacterium. the outer membrane protein 25 kda (omp25) gene plays an important role in simulating of tnf-α, ifn-α, macrophage, and cytokines cells. in the current study molecular cloning and expression analysis of omp25 gene for designing a subunit vaccine against brucella was investigated. amplifying the full length of candidate gene was performed using specific primers. sub-cloning of this gene conducted using ptz57r/t vector in top10f`strain of escherichia coli(e.coli) as the host. also, pet32(a) + vector used for expression in bl21 (de3) strain of e.coli . omp25 gene with 642 bp size was amplified and cloned successfully. the expression results were confirmed by sequencing and sodium dodecyl sulfate polyacrylamide gel electrophoresis (sds-page) analyses which showed 42 kda protein band correctly. also, phylogenic analysis showed this gene has a near genetic relation with other brucella strains. according to our results we can propose this gene as a candidate useful for stimulation of cell-mediated and humoral immunity system in future study.