Cloning of outer membrane protein gene Omp31 Brucella melitensis

Brucellosis is one of the most important zoonotic diseases transmitted to humans from animals or animal products. Brucella melitensis is the most pathogenic species in the genus brucella. The outer membrane protein 31 (Omp31) of B. melitensis is considered to be a protective immunogen and an important candidate vaccine. In this study, cloning from B. melitensis Rev 1 strain of Omp31 coding gene (omp31) was aimed. Brucella melitensis REV-1 live vaccine strain was used in the study. After reactivation of strain, DNA was extracted from bacterial culture. Gene sequence which encodes outer membrane protein was obtained from Gene Bank. Primers were designed to clone this region. After primer design, the gene region amplified by the polymerase chain reaction was cloned into the pET28a expression vector and transformed into Escherichia coli BL21 bacteria. Plasmid extraction was performed from E. coli BL21. The insert and plasmid were separately observed as a result of cutting with the restriction enzyme used for cloning. Expected size (790 bp) insert amplified with cloning primers again and amplicon was sequenced. The sequence obtained after sequencing analysis was compared to the gene bank and confirmed to be the outer membrane protein of B. melitensis. Further studies are required to investigate the antigenic properties of the cloned recombinant outer membrane protein Omp31 (rOmp31) and determine the potential to be a candidate for vaccination