奶牛温氏支原体16S rRNA基因的PCR扩增、克隆及分析  

SEQUENCING AND PHYLOGENETIC ANALYSIS OF THE 16S RRNA GENE OF MYCOPLASMA WENYONII-CGXD STRAIN

作　　者：苏乾莲[1] 黄维义[1] 张为宇[1] 陈明[2] 拉伊萨[1] 

机构地区：[1]广西大学动物科学技术学院,南宁530005 [2]广西水产研究所,南宁530022

出　　处：《中国兽医寄生虫病》2008年第6期1-5,共5页Chinese Journal of Veterinary Parasitology

基　　金：广西大学科研基金项目(X032066)

摘　　要：目的对奶牛温氏支原体16S rRNA基因进行PCR扩增及克隆分析。方法从自然感染体的广西奶牛无菌采集血液,分离温氏支原体并提取病原基因组,用血营养菌的16S rRNA基因的通用引物进行PCR扩增,将扩增产物克隆到PGEM-Teasy载体后进行测序和分析,并与Gene bank上搜索的温氏支原体相应序列进行比较,建立系统发育树。结果PCR扩增得到长约1.5 kb的扩增片段,测序结果显示该片段全长为1 453 bp,同源性分析表明该序列与Neimark公布的温氏支原体(前称温氏附红细胞体)(AF016546)的16S rRNA基因序列同源性达到97.4%,与系统发育进化树表明本株温氏支原体同本地株的关系较近,而与国外株的新缘关系较远。国内公布的广西株同源性为99.8%。结论结果表明证实该病原为温氏支原体,从分子生物学水平证实了温氏支原体在广西的存在。由于本试验分离得到的牛温氏支原体与国外发表的牛温氏支原体核苷酸序列相差2.6%,因此两者的基因型存在一定的差异,这对该病的分子流行病学分析具有一定的意义。The complete 16S rRNA gene of Mycoplasma. wenyoni i-CGXD strain was determined by PCR. It was sequenced and blasted. The results showed that the sequence was 97.4% identical to that of Neimark published sequence （AF016546） The 1453 bp complete sequence was compared with the sequence of mycoplasmas available in GenBank. The data were analyzed and a phylogenetic tree was established. The results indicated that the hereditary distance of M. wenyonii was close to local strain, and the hereditary distance was far from foreign, strain. The result indicated that there were some genetic differences between different isolated strains in M wenyonii. These findings should have important implications for studying of molecular epidemiology and the diagnosis of M wenyonii.

关 键 词：温氏附红细胞体 16S RRNA基因 克隆 序列分析 

分 类 号：S945.262[农业科学—水产养殖] Q78[农业科学—水产科学]