|
|||||
|
|
| 牛白血病病毒GP51蛋白在大肠杆菌中表达及胶体金免疫层析试纸条检测方法的建立 | |
| 引用本文： | 宋阳威,张鑫宇,张常印,吴亚力,孙怀昌,唐泰山,张敬友.牛白血病病毒GP51蛋白在大肠杆菌中表达及胶体金免疫层析试纸条检测方法的建立[J].中国预防兽医学报,2010,32(4). |
| 作者姓名： | 宋阳威 张鑫宇 张常印 吴亚力 孙怀昌 唐泰山 张敬友 |
| 作者单位： | 1. 徐州出入境检验检疫局,江苏,徐州,221006 2. 扬州大学,兽医学院,江苏,扬州,225009 3. 江苏出入境检验检疫局,江苏,南京,210001 |
| 基金项目： | 江苏检验检疫局科研项目 |
| 摘 要： | 为了建立一种快速诊断牛白血病(BLV)的检测方法,本研究从BLV中扩增出其囊膜糖蛋白gp51基因,经测序后克隆于表达载体pET32a(+)中,构建了重组表达质粒pET32a-gp51。将其转化受体菌BL21,用IPTG诱导后表达出约42ku的目的蛋白,经western blot检测表明目的蛋白具良好的反应原性。以胶体金标记的羊抗牛IgG(Fc)抗体作为标记抗体,将纯化的His-gp51重组蛋白和羊抗牛IgG分别标记于硝酸纤维素膜上作为检测线和质控线,各部件按序装配形成快速诊断试纸条。对22份样本分别用试纸条和ELISA试验进行检测,2种方法的阳性符合率为88.9%(8/9),表明本研究建立的胶体金免疫层析方法简便、快捷,具有较好的特异性和一定的敏感性,适宜基层初筛诊断和现场应用。 |
| 关 键 词： | 牛白血病病毒 gp51蛋白 表达 胶体金 免疫层析法 |
Expression of bovine leukemia virus GP51 protein in E.coli and development of colloidal gold immunochromatographic diagnosis strips for detection of antibody against the virus |
|
| SONG Yang-wei,ZHANG Xin-yu,ZHANG Chang-yin,WU Ya-li,SUN Huai-chang,TANG Tai-shan,ZHANG Jing-you.Expression of bovine leukemia virus GP51 protein in E.coli and development of colloidal gold immunochromatographic diagnosis strips for detection of antibody against the virus[J].Chinese Journal of Preventive Veterinary Medicine,2010,32(4). | |
| Authors: | SONG Yang-wei ZHANG Xin-yu ZHANG Chang-yin WU Ya-li SUN Huai-chang TANG Tai-shan ZHANG Jing-you |
| Institution: | SONG Yang-wei1,ZHANG Xin-yu2,ZHANG Chang-yin3,WU Ya-li1,SUN Huai-chang2,TANG Tai-shan3,ZHANG Jing-you3(1. Xuzhou Entry-Exit Inspection , Quarantine Bureau,Xuzhou 221006,China,2. College of Veterinary Medicine,Yangzhou University,Yangzhou 225009,3. Jiangsu Entry-Exit Inspection , Quarantine Bureau,Nanjing 210001,China) |
| Abstract: | The gp51 gene of BLV was amplified from the bovine leukaemia virus(BLV) ,cloned into the expression vector pET32a(+) ,and expressed in E.coli BL21 cells. The recombinant protein was detected by western blot and showed similar immuno-reactiongenicity to native GP51. A rapid gold immumochromatographic(GICA) strip was assembled by spraying the purified recombinant protein on the nitrocellulose membrane as the test line(T line) ,and the Gold-conjugated goat anti-bovine IgG as the control line(C line) . Test on ... |
| Keywords: | bovine leukaemia virus gp51 protein expression colloid gold immunochromatographic assay |
| 本文献已被 CNKI 万方数据 等数据库收录！ | |