Construction of an Expression Plasmid (Vector) Encoding Brucella melitensis Outer Membrane Protein, a Candidate for DNA Vaccine

Background: DNA vaccination with plasmid encoding bacterial, viral, and parasitic immunogens has been shown to be an attractive method to induce efficient immune responses. Bacteria of the genus Brucella are facultative intracellular pathogens for which new and efficient vaccines are needed. Methods: To evaluate the use of a DNA immunization strategy for protection against brucellosis, a plasmid containing the DNA encoding the Brucella melitensis (B. melitensis) ۳۱ kDa outer membrane protein, as a potent immunogenic target, was constructed. Results: The constructed plasmid, pcDNA۳.۱+omp۳۱, was injected intramuscularly into mice and the expression of omp۳۱ RNA was assessed by RT-PCR. The integrity of the pcDNA۳.۱+omp۳۱ construct was confirmed with restriction analysis and sequencing. Omp۳۱ mRNA expression was verified by RT-PCR. Conclusion: Our results indicate that the pcDNA۳.۱+omp۳۱ eukaryotic expression vector expresses omp۳۱ mRNA and could be useful as a vaccine candidate.

Keywords:

Brucella melitensis , DNA Vaccine , Omp۳۱ , PcDNA۳.۱