宁夏布鲁氏菌临床分离株OMP25基因序列特征分析.docVIP

宁夏布鲁氏菌临床分离株OMP25基因序列特征分析* 马学平，高建炜，詹军，李丽 （宁夏疾病预防控制中心，宁夏银川，750004） 摘要：目的 鉴定宁夏布鲁氏菌临床分离株，扩增OMP25基因并测序，测序结果与Genebank公布的布鲁氏菌6个种的核苷酸和推导氨基酸序列比对，分析宁夏地方株OMP25基因序列特征，为布鲁氏菌鉴定以及后续的基因克隆与表达研究提供基础。方法 参考Genebank公布的布鲁氏菌外膜蛋白OMP25基因序列设计1对引物，采用聚合酶链式反应（PCR）从布鲁氏菌扩增基因组中扩增omp25基因，经电泳检测后送基因公司双向测序，测序结果用Contig Express软件拼接，DNAstar软件分析。结果 经鉴定，分离2株布鲁氏菌为羊种3型，测序获得OMP25基因序列全长序列为638bp，编码212个氨基酸，核苷酸序列与Genebank公布羊种布鲁氏菌OMP25基因同源性为99.9%，氨基酸序列同源性为99.7%。系统进化树上显示，分离株与参考布鲁氏菌亲缘关系相近，与羊种布鲁氏菌为同一个分枝。结论 宁夏布鲁氏菌分离株OMP25基因序列与参考菌株序列同源性很高，且与羊种布鲁氏菌在同一分枝，说明OMP25基因序列是一个保守序列，为布鲁氏菌的分子生物学鉴定和诊断试剂以及疫苗研制提供基础。 关键词：布鲁氏菌，OMP25基因，序列特征分析 Analysis OMP25 gene sequence characteristics of clinical isolates Brucella from Ningxia MA Xue-ping,GAO Jian-wei, ZHAN Jun, LI li (Ningxia Center for Disease Prevention and Control, Yinchuan 750004, China) 【Abstract】Objective To identify clinical isolates Brucella and amplify OMP25 gene sequencing, the results of sequencing Genebank published Brucella 6 kinds of nucleotide and deduced amino acid sequence alignment, analysis OMP25 gene sequences characteristics of isolates from Ningxia, in order to provide basis for Brucella identification and gene cloning and expression studies. Methods Brucella OMP25 gene sequence designed one pairs of primers by reference Genebank, polymerase chain reaction (PCR) amplify omp25 gene from the genome of Brucella, gene fragments were detected by agarose gel electrophoresis and the sequencings of PCR products, the results of sequencing using Contig Express software mosaic and analysis by DNAstar software. Results The brucella strains was identified as type 3 of B.melitensis，and get full-length sequence of OMP25 gene sequence of 638bp by sequencing, and encodes 253 amino acid, the nucleotide sequence with the published Genebank of B.melitensis OMP25 gene homology is 99.9%, the amino acid sequence homology is 99.7%, Display the phylogenetic tree, strain and reference of Brucella separation of close relationship, and B.melitensis is a branch of *【基金项目】宁夏自然科学基金项目（NZ10201