Sequence Analysis Of Some Functionality Genes Of Brucella Vaccines And Study On A New Diagonostic Technique AMOS-PCR

In this study,we studied one more advanced PCR diagonosis technique AMOS-PCR for brucellosis,which can identifified the biotypes of brucella,by using it to identifify brucella vaccines B.Melitensis M5,B.abortus 104M,B.abortus S19 and B.suis S2 in China.And at the same time using Primers in the method to amplified the aingle gene of each biotype of brucella.sp.Secondly we get some published Primers,using them to squence and analysis five protection antigen genes Omp31,Omp25,Omp2b,p39,L7/L12 and a virulence gene virb8 from Type IV secretion systems(TFSS) of brucella.Result (1)The AMOS-PCR has do a great job in identification of brucella.spp.and the single AMOS-PCR also showed that the method has great specific among brucella Abortus,Melitensis,Suis.(2) Successfully amplified the five protection antigen genes and get the sequence of each vaccine strains all five genes. L7/L12 get 375bp;p39 get 1207bp;Omp2bget 1089bp;Omp31 get 723bp;Omp25 get 642bp.Sequence analysis shows that among all the genes,S2,M5 used in China share a great homological to those abroad, homological as high as 97.1%～100% for S2,and 95.1%～100% for M5.Among all the genes we sequence ,the Omp2b gene shows more notable distance in homological. S2 Omp2b with others and M5 Omp2b share a 97.1～98% homological.(3) virb8 gene of 7 brucella vaccine strains (S19,104M,S2,JYS2, M5,TKM5,42#) were Sequenced,the virb8 was 720bp. Sequence analysis shows that among all 7 strains of brucella,the virb8 gene share great homological as high as 99.7%～100%.