中国兽医科学2013，Vol.43Issue(7)：723-727,5.
鸭疫里氏杆菌外膜微孔蛋白基因的克隆和序列分析及PCR检测方法的建立
Cloning and sequence analysis of outer membrane phosphate selective Porin protein gene of Riemerella anatipestifer and development of PCR detection method
摘要
Abstract
Specific primers were designed according to outer membrane phosphate selective Porin protein gene sequence of Riemerella anatipestifer available in GenBank.Target fragments of 1 212 bp were successfully cloned from genomic DNA of serotypes 1,2,6,10,11,13,14 and 17 R.anatipestifer strains.Multiple sequence alignment analysis showed the homology of the porin gene between the 8 serotype strains was more than 99.6％.Specific primers were designed for 357 bp fragment to establish a PCR diagnostic method.The 357 bp target fragment was not amplified from genome of duck Pasteurella,duck pathogenic Escherichia coli,Salmonella Typhimurium and duck hepatitis virus,which indicated that the method had a high specificity.The lowest amount of DNA of R.anatipestifer detectable by this method was 73.96 pg.Sharp interest bands with a size of 357 bp were amplified from 8 isolates of R.anatipestifer of Jilin,Hebei and Beijing.Coincidence with identification of bacteria was 100％.Result provides identification method for R.anatipestifer and a theoretical basis for subsequent research of Porin.关键词
鸭疫里氏杆菌/微孔蛋白/聚合酶链式反应/检测方法Key words
Riemerella anatipestifer / Porin/ PCR / detection method分类
农业科技引用本文复制引用
么乃全,王全凯,姜秀云,徐凤宇,于丹,孟轲音,高云航..鸭疫里氏杆菌外膜微孔蛋白基因的克隆和序列分析及PCR检测方法的建立[J].中国兽医科学,2013,43(7):723-727,5.基金项目
吉林省教育厅"十二五"科学技术研究项目(201372) （201372）
吉林省科技发展计划项目(20110221) （20110221）
国家现代农业产业技术体系建设专项(CARS-43) （CARS-43）
国家星火计划项目(2012GA660003) （2012GA660003）
