Clonging And Sequence Analysis Of Partial Outer Membrane Protein Genes Of Brucella Strain Isolated In Inner Mongolia

The primers were designed according to Brucella gene sequence of GenBank. The OMP19, OMP28, OMP25 and OMP31 genes of NH-B1, NH-B2, NH-B3, NH-R1, NH-R2 and NH-R3 isolated from Inner Mongolia were amplified by PCR and purified. The PCR product were ligated with PMD19-T vector, transformed into DH5αcells,extracted plasmid,the recombinant plasmid obtained were identified by PCR and sequenced. The results showed that the length of OMP19 gene was 618bp, which include a whole open reading frame and located in 43～576bp; The length of OMP28 gene was 953bp, which include a whole open reading frame and located in 151～903bp; The length of OMP25 gene was 865bp and located in 166～808 bp; The length of OMP31 gene was 1167bp and located in 137～859bp.The nucleotide sequences homology was compared with reference Brucella gene sequence by DNAstar software. The results showed that the homology of OMP19 nucleotide sequences was ranged from 99.8% to 100% among the 6 Brucella isolates and 99.4% to 100% compared with reference Brucella gene sequence; the homology of OMP28 nucleotide sequences were ranged from 99.9% to 100% among the 6 Brucella isolates (the homology of NH-B2,NH-B3,NH-R1 and NH-R2 were 100%, the same as NH-B1 and NH-R3; the homology of NH-B2,NH-B3,NH-R1,NH-R2 and NH-B1, NH-R3 were 99.9%) and 99.6% to 100% compared with reference Brucella gene sequence. The homology of OMP25 nucleotide sequences were ranged from 99.5% to 100% among the 6 Brucella isolates (the homology of NH-B2,NH-B3,NH-R1, NH-R2 and Brucella abortus S19 were 100%, the same as NH-B1 and Brucella suis 1330; the homology of NH-B2,NH-B3,NH-R1,NH-R2 and NH-B1, NH-R3 were 99.9%)and 99.6% to 100% compared with reference Brucella gene sequence. The homology of OMP31 nucleotide sequences, which were in NH-B1, NH-B2 and NH-R3, were 100%, which was ranged from 99.6% to 100%. The homology among the 6 Brucella isolates were above 99% and the homology of the 6 Brucella isolates and reference Brucella gene sequence were above 98%. This paper indicates that the OMP19, OMP25, OMP28 and OMP31 were consistent, the same as consulted literature report.