Cloning of outer membran protein gene of brucella melitensis

In this study cloning of Brucella melitensis outer membrane protein was aimed. It was thought that, this study could be a potential and national preliminary work which would lead a way for rapid diagnosis kit and vaccine development. Brucella melitensis REV-1 live vaccine strain was used in the study. After isolation and confirmation of strain, DNA was extracted from bacterial culture. Gene sequence which encodes outer membrane protein was obtained from Gene Bank and primers for gene were designated. PCR reaction was carried out and amplicon was cloned Pet28A expression vector, then recombinant plasmid was transformed into Escherichia coli BL21. Plasmid was extracted from E. coli BL21 strain and after the restriction digest insert and plasmid were observed separately. Insert with size of 790 bp, amplified with cloning primers again and amplicon was sequenced. Obtained sequence was compared with those in Genbank and insert was confirmed as B. melitensis outer membrane protein. Recombinant outer membrane protein Omp31 (rOmp31) has a feature of experimental component for vaccine candidate researches. In this way this study could be a potential preliminary national work for further studies such as developing rapid diagnostic kits.