Isolation And Cloning Of AapJ Gene From Brucella melitensis

Economic losses resulting from the impact of brucellosis on animals have increased, which is reflected in turn on humans. Brucella is pathogenic bacteria shared by humans and pet animals. They still pose a significant risk that threatens health and economics in Syria. Many efforts have focused to prevent the spread of brucellosis disease, by resorting to the use of vaccines prepared from Brucella strains, where now the target is preparation of vaccine components proteins of Brucella can be used as a vaccine or an improved traditional vaccine. In this study, the aapJ gene encoding permeas amino acid has been isolated and amplified by Polymerase chain reaction (PCR) using B. melitensis16M genomic DNA, and using aqualitative primer containing restriction sites of both XhoI-BamHI. It was found that the length of the segment was 1032 pb. Then, the segment was inserted to the plasmid gene expression (pET-15b) that was cut by the same restriction enzymes, then the recombinant plasmid was put into the E. coli DH10B bacteria, by bacterial transformation technology, so as to clone the gene, the next step was to choose colonies that grew on media containing ampicillin, extract the plasmid of them, and treat it by the two restriction enzymes Xhol and BamHI, so as to confirm that it contains the aapJ gene cloned gene sequence analysis revealed, no mutations were introduced during the cloning process. Key world: B. melitensis, aapJ, PCR, Isolated, Cloning, Sequence.