山羊奇异变形杆菌的分离鉴定及16S rDNA和ZapA基因的PCR-RFLP分析  

Isolation and identification of Proteus mirabilis from goat and the analysis of 16S rDNA and ZapA gene by PCR-RFLP

作　　者：王豪举[1] 倪莉[1] 杨红军[2] 赵俊[1] 徐宗可[3] 徐强[1] 徐丽敏[1] 

机构地区：[1]西南大学动物科技学院重庆市牧草与草食家畜重点实验室,重庆400715 [2]西南大学资源环境学院,重庆400715 [3]中国农业科学院兰州兽医研究所,甘肃兰州730046

出　　处：《中国兽医学报》2011年第11期1594-1598,共5页Chinese Journal of Veterinary Science

基　　金：重庆市教委重点项目(kj070204)

摘　　要：从发病山羊分离到5株病原菌,对分离株进行游散行为分析,生化试验、动物试验、药敏试验及16SrDNA和ZapA基因克隆测序,选用限制性内切酶对分离株16SrDNA和ZapA基因进行PCR-RFLP分析。结果表明分离株出现游散生长现象,对丁胺卡那霉素和复达欣敏感,小鼠LD50为2.500 0×107 CFU/mL～4.445 3×107 CFU/mL;分离株16SrDNA与奇异变形杆菌的同源率为99.5%～99.8%,ZapA基因与奇异变形杆菌的同源率为99.5%;PCR-RFLP分析发现,分离株酶切片段数目和大小与标准株相同。结果表明分离株是奇异变形杆菌,多态性较为单一。Five strains of pathogen were isolated from the infected goats. Swarming behaviour assays, biochemical, animal and antibiotic resistance tests were carried out to identify its properties. Then 16S rDNA and ZapA were cloned and sequenced and both 16S rDNA and ZapA were analyzed with PCR-PFLP after digestion by specific restriction enzymes. The data showed the field strains had the swarming behaviour,which were sensitive to amikacin and for- turn,the LDs0 in mice was from 2. 500 0× 10^7 CFU/mL to 4. 445 3× 10^7CFU/mL. The DNA sequencing showed 99. 5%-99.8% similarity with Proteus mirabilis for 16S rDNA and 95. 5% for ZapA gene; the segments from PCRRFLP analysis displayed no disparity with reference strain in number and size. The results indicate that the field strains are a member of Proteus rnirabilis with no polymorphism.

关 键 词：奇异变形杆菌 分离 鉴定 PCR—RFLP 

分 类 号：S852.61[农业科学—基础兽医学]