牛腐蹄病坏死杆菌H05菌株白细胞毒素基因的克隆与序列分析  

Clone and Sequence Analysis of Leukotoxin Gene of Fusobacterium Necrophorum H05 Strain Isolated From Cattle

作　　者：郭东华[1] 孙玉国 李林[3] 王君伟[3] 张建涛[3] 王洪斌[3] 

机构地区：[1]黑龙江八一农垦大学动物科技学院 [2]黑河市畜牧兽医局疾病预防与控制中心 [3]东北农业大学动物医学院

出　　处：《黑龙江八一农垦大学学报》2008年第4期48-51,共4页journal of heilongjiang bayi agricultural university

基　　金：国家科技攻关项目(2002BA518A04)

摘　　要：以黑龙江省分离的牛腐蹄病坏死杆菌H05菌株基因组DNA为模板,参考GenBank发表的羊腐蹄病坏死杆菌A25菌株白细胞毒素基因核苷酸序列,设计5对覆盖白细胞毒素基因完整开放阅读框(ORF)的引物,通过PCR扩增获得了H05菌株白细胞毒素基因5个相互重叠的DNA片段,将它们克隆到pMD18-T载体中。测序结果显示:牛腐蹄病坏死杆菌H05菌株白细胞毒素基因长为9723bp,编码一个3240个氨基酸的蛋白质,与羊腐蹄病坏死杆菌白细胞毒素蛋白的核苷酸和氨基酸的同源性分别为99.75%和99.63%,缺失了1078位的天冬酰胺。Five pair of primers was designed to amplify five overlapping fragments of leukotoxin gene .of Fusobacterium necrophorum H05 strain isolated from cattle of Heilongjiang province by PCR based on nucleotide sequences of leukotoxin gene of Fusobacterium necrophorum A25 strain isolated from goat in GenBank. Five overlapping fragments of leukotoxin gene of Fusobacterium necrophorum H05 strain was cloned to pMD18-T vector. These inserts were sequenced and the results showed that leukotoxin gene of Fusobacterium necrophorum H05 strain consisted of 9,723 base pairs, encoded 3,240 amino acides. Identities of nucleotides and amino acides compared with leukotoxin gene of Fusobacterium necrophorum A25 strain were respectively 99.75 % and 99.63 %, with deletion of aspartic acid at position of 1078 amino acid. Molecular cloning of leukotoxin gene of Fusobacterium necrophorum H05 strain obtained lay the foundation for research on development of footrot vaccine and a diagnostic kit applying to detection of footrot.

关 键 词：牛腐蹄病 坏死杆菌 白细胞毒素基因 序列分析 

分 类 号：S858.23[农业科学—临床兽医学] Q785[农业科学—兽医学]