河南及山西省21株鸡杆菌分离株16S rRNA基因序列分析

Sequence analysis of 16S rRNA genes of twenty-one Gallibacterium isolates from Henan and Shanxi provinces

对21株分离自河南及山西地区的鸡杆菌的16SrRNA基因序列进行研究,以此分析分离株的进化关系并确定其在鸡杆菌属中所属的具体种。采用PCR方法扩增其16SrRNA基因序列,然后克隆测序。用DNAStar软件对21个菌株的序列及GenBank中已公布的鸡杆菌属相关菌株序列进行同源性比较,结果发现21株分离株之间的核苷酸序列同源性为96.0%～100%,同鸭源鸡杆菌F279(Gallibacterium anatis F279)(AF228002)的同源性为95.3%～99.3%,同输卵管炎鸡杆菌F150(Gallibacterium salpingitidis F150)(EU424000)的同源性为88.3%～91.5%,同虎皮鹦鹉鸡杆菌F450(Gallibacterium melopsittaci F450)(EU339196)的同源性为90.3%～93.3%,同海藻糖发酵鸡杆菌52-S3-90(Gallibacterium trehalosifermentans 52-S3-90)(EU339199)的同源性为88.2%～91.4%,同多杀性巴氏杆菌CCUG 179(P.multocida CCUG 17977)(AF294411)的同源性为85.5%～88.8%,对21株细菌的序列同以上菌株的16SrRNA基因序列进行遗传进化树分析,显示21株细菌同鸭源鸡杆菌F279(AF228002)构成一个单独的大分支。结果表明,所有分离株均属于鸭源鸡杆菌,首次大量报道了来源于河南、山西省不同地区的鸡杆菌的16SrRNA基因序列,为鸡杆菌分类的研究提供了参考。

To analyse the evolutionary relationship of the twenty-one isolates of Gallibacterium from Henan and Shanxi provinces and identify the species of them, 16S rRNA gene were amplified by PCR and the nucleotide se- quences were determined. All nucleotide sequences of the strains which were from different chicken flocks were compared by software of DNAStar with the related 16S rRNA gene sequences of Gallibacterium in GenBank. Results showed the homology of nucleotide sequences were 96.0%-100~ among twenty-one isolates,and the homology of twenty-one isolates with Gallibacterium anatis F279（AF228002）was 95.3 ^-99.3 %, while the homologies of twen- ty-one isolates with Gallibacterium salpingitidis F150（EU424000） ,Gallibacterium melopsittaci F450（EU339196）, Gallibacterium trehalosifermentans 52-S3-90 （EU339199） and P. multocida CCI.JG 17977 （AF294411） were 88. 3%- 91.5% ,90. 3%-93.3% ,88. 2%-91.4% and 85.5%-88.8% ,respectively. Phylogenetic tree showed that the twenty-one i- solates and the strain Gallibacteriurn anatis F279 fell into the same branch,all the isolates were classified in Gallibacterium anatis. A number of 16S rRNA gene sequences of Gallibacterium from different areas in Hcnan and Shanxi provinces in this research is firstly reported in China,it will provide a reference for further study in classification of Gallibacterium.