奶牛乳腺炎大肠杆菌csgC基因的克隆及序列分析  

Cloning and Sequencing Analysis of Escherichia coli Dairy Cattle Mastitis Strain csgC Gene

作　　者：王林锋[1,2] 杨宏军[3] 杨少华[3] 王长法[3] 高运东[3] 仲跻峰[3] 葛利江[1] 

机构地区：[1]山东农业大学动物科技学院 [2]山东省农业科学院奶牛研究中心,济南250100 [3]山东省农业科学院奶牛研究中心

出　　处：《生物技术通报》2008年第5期163-165,170,共4页Biotechnology Bulletin

基　　金：863项目;山东省农科院重大成果培育基金(2006YCG012);山东省农科院高技术自主创新基金(2006YCX027)

摘　　要：根据GenBank上发表的curli菌毛csgC基因序列,设计了一对特异性引物。从患乳腺炎的奶牛乳汁中分离出致病性大肠杆菌,经生物学鉴定后,提取全基因组DNA为模板,PCR扩增出csgC基因,连入pMD18-T克隆载体,测序。结果表明,扩增片段含有333个核苷酸,编码111个氨基酸的成熟蛋白,与已报道的大肠杆菌W3110的全基因组DNA中的csgC基因序列最相近,氨基酸序列同源性为99.7%。Curli菌毛csgC基因的克隆,为获得重组csgC蛋白及对其结构和功能的研究奠定了基础。One pair of specific primers to curli fibers csgC gene was designed and synthesized based on the published sequence in GenBank. Escherichia coli were isolated and identi fled from the milk of cows with mastitis, csgC gene was cloned from genomic DNA by PCR,the product of which was inserted into vector pMD18-T to construct plasmid pMD18-T/ csgC. Sequencing analysis showed that the amplified fragment was composed of 333 nucleofides, which encode a mature polypeptide with 111 amino acids mature protein. Compared with the reported csgC gene from whole genome DNA sequence of W3110,the highest amino acid identity was 99.7%. The results showed that the cloning plasmid was constructed successfully,thus laid a foundation for the structure and function of csgC protein.

关 键 词：大肠杆菌 curli菌毛 csgC基因 

分 类 号：S852.61[农业科学—基础兽医学] Q78[农业科学—兽医学]