山羊源海藻糖比伯斯坦杆菌(Bibersteinia trehalosi)转铁结合蛋白B的克隆与功能鉴定  

Expression and specific identification of transferrin binding protein B in Bibersteinia trehalosi

作　　者：郭建华 焦云娟 霍溢徽 段杨鑫婷 何深宏 刘威[1] 任绍科 罗献梅 虞荣华 程方俊[1] GUO Jian-hua;JIAO Yun-juan;HUO Yi-hui;DUAN Yang-xin-ting;HE Shen-hong;LIU-Wei;REN Shao-ke;LUO Xian-mei;YU Rong-hua;CHENG Fang-jun(College of Animal Science,Southwest University,Chongqing 402460,China;Cumming School of Medicine,University of Calgary,Alberta T2N 4N1,Canada)

机构地区：[1]西南大学动物科学学院,重庆荣昌402460 [2]加拿大卡尔加里大学医学院,阿尔伯塔卡尔加里T2N 4N1

出　　处：《中国兽医学报》2020年第1期97-101,共5页Chinese Journal of Veterinary Science

基　　金：重庆市社会民生科技创新专项资助项目(cstc-2016shmszx0932)

摘　　要：山羊海藻糖比伯斯坦杆菌(Bibersteinia trehalosi)是造成山羊肺炎重要病原菌之一,其转铁结合蛋白B(transferrin binding protein B,TbpB)结构与功能的研究对于阐明其致病机理有重要作用。本研究在前期分离鉴定菌株基础上,通过生物信息学分析结果显示,海藻糖比伯斯坦杆菌TbpB包含氨基小叶(N-lobe)和羧基小叶(C-lobe)。分别对该菌TbpB^ 20-581 (In-tact)、N-lobe、C-lobe进行克隆、表达与纯化,对纯化的蛋白与山羊转铁蛋白、绵羊转铁蛋白、猪转铁蛋白、牛转铁蛋白、鸡转铁蛋白进行特异性点阵杂交反应,发现山羊海藻糖比伯斯坦杆菌TbpB In-tact以及N-lobe与山羊转铁蛋白、绵羊转铁蛋白有特异结合反应,而与猪转铁蛋白、鸡转铁蛋白没有反应,山羊海藻糖比伯斯坦杆菌TbpB C-lobe与山羊转铁蛋白、绵羊转铁蛋白、猪转铁蛋白、牛转铁蛋白、鸡转铁蛋白均无反应。结果表明山羊海藻糖比伯斯坦杆菌TbpB与山羊转铁蛋白结合发生在N-lobe。In-tact,N-lobe and C-lobe of transferrin binding protein B from Bibersteinia trehalosi were cloned,expressed and purified in the experiment.The purified proteins were conducted binding experiment to HRP-goat transferrin,sheep transferrin,porcine transferrin,bovine transferrin and chicken transferrin,respectively.The results showed that In-tact and N-lobe TbpB from Bibersteinia trehalosi could bind to goat-transferrin and sheep-transferrin but could not bind to porcine transferrin and chicken transferrin.C-lobe of TbpB from Bibersteinia trehalosi could not bind to either transferrin.The binding site between TbpB and goat transferrin is in N-lobe.

关 键 词：山羊 海藻糖比伯斯坦杆菌 转铁结合蛋白B 

分 类 号：S852.61[农业科学—基础兽医学]