摘要
目的目的对宁夏布鲁氏菌Omp31基因序列进行分析,为菌株种型鉴定和试剂研究奠定基础。方法根据Genebank公布Omp31基因序列设计引物,采用PCR扩增、电泳检测、胶回收、纯化、送基因公司双向测序,用Contig Express软件对测序结果拼接,用DNAstar软件进行结果分析。结果经PCR扩增测序得到Omp31基因全长序列大小为723 bp,编码240个氨基酸,与Genebank上羊种布鲁氏菌Omp31基因同源性达99.9%,氨基酸序列的同源性100%。通过遗传分析,分离株与参考菌亲缘关系相近,与羊种布鲁氏菌处在同一分枝。结论宁夏布鲁氏菌Omp31序列与发布的参考序列之间的同源性很高,而且与羊种布鲁氏菌处在同一分支。
Objective To analyze Omp31 gene sequences characteristic of Brucella,which isolate from Ningxia,in order to pro- vide basis for Brucella identification and gene cloning and expression. Methods Brueella Omp31 gene sequences were designed one pairs of primers by reference Genebank,polymerase chain reaction （PCR） amplify Omp31 gene from the genome of Brucella,gene frag- ments were detected by agarose gel electrophoresis and the sequences of PCR products. Use the Contig Express software to joint the result of sequencing. Results The brucella strains was amplified, and get full - length sequence of Omp31 gene sequence of 723 bp by sequen- cing,and encodes 240 amino acid, the nucleotide sequence with the published Genebank of B. melitensis Omp31 gene homology was 99.9%, the amino acid sequence homology was 100%, Display the phylogenetic tree, strains of references of Brucella and separations had close relationship, and at the same branch with B. melitensis. Conclusion The Omp31 gene of brucella is isolated strains and reference strains has high homology and in the same branch of B. melitensis. Omp31 gene is a conserved sequence, the research results provide the basis for molecular biology identification and subsequent diagnostic reagent.
出处
《宁夏医学杂志》
CAS
2015年第11期967-969,共3页
Ningxia Medical Journal
基金
宁夏自然科学基金资助项目(NZ1296
NZ14225)
宁夏卫生厅重点科研计划项目(2012080)
