| 艰难梭菌A毒素受体结合区基因在乳链菌中的表达 |
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| 引用本文： | 杨晓强,赵亚刚,孙大勇,姜泊,陈学清.艰难梭菌A毒素受体结合区基因在乳链菌中的表达[J].第四军医大学学报,2009(18):1676-1680. |
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| 作者姓名： | 杨晓强 赵亚刚 孙大勇 姜泊 陈学清 |
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| 作者单位： | 广州军区广州总医院消化内科;南方医科大学附属南方医院消化内科;广州医学院第一附属医院消化内科; |
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| 基金项目： | 国家自然科学基金自由申请面上项目(30570839); 广东省自然科学基金(05004735) |
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| 摘 要： | 目的：在乳链菌内表达艰难梭菌A毒素受体结合区并检测其活性．方法：提取艰难梭菌标准株基因组DNA，扩增编码艰难梭菌毒素A毒素受体结合区的羧基未端基因重复序列（14CDTA），分别连接到乳链菌表达载体pNBC1000及pNBC2000，转化乳链菌，蛋白电泳及Western—blot检测蛋白定位表达情况．结果：成功构建了14CDTA乳链菌表达载体，并在乳链菌内表达了艰难梭菌受体结合区蛋白，分泌表达的艰难梭菌受体结合区蛋白大小约39．2ku，膜锚定表达的受体结合区蛋白大小约为55．1ku，所表达的蛋白均可与艰难梭菌A毒素多克隆抗体发生免疫印迹反应．结论：表达艰难梭菌受体结合区蛋白的重组乳链菌可以识别艰难梭菌A毒素多克隆抗体，为研制防治艰难梭菌疫苗的口服生态活菌亚单位疫苗奠定了一定基础．
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| 关 键 词： | 艰难梭菌 A毒素 受体结合区 乳链菌 |
Expression of Clostridium difficile toxin A receptor binding domain in Lactococcus lactis |
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| YANG Xiao-Qiang,ZHAO Ya-Gang,SUN Da-Yong,JIANG Bo,CHEN Xue-Qing.Expression of Clostridium difficile toxin A receptor binding domain in Lactococcus lactis[J].Journal of the Fourth Military Medical University,2009(18):1676-1680. |
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| Authors: | YANG Xiao-Qiang ZHAO Ya-Gang SUN Da-Yong JIANG Bo CHEN Xue-Qing |
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| Institution: | YANG Xiao-Qiang1,ZHAO Ya-Gang1,SUN Da-Yong1,JIANG Bo2,CHEN Xue-Qing31Department of Gastroenterology,Guangzhou General Hospital of Guangzhou Command,Guangzhou 510010,China,2Department of Gastroenterology,Nanfang Hospital,Southern Medical University,Guangzhou 510515,3Department of Gastroenterology,First Affiliated Hospital,Guangzhou Medical College,Guangzhou 510120 |
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| Abstract: | AIM: To expreess the Clostridium difficile toxin A receptor binding domain and test its bioactivity.METHODS: The gene of fourteen of the 38 C-terminal toxin A repeats(14CDTA) coding for the clostridium difficile toxin A receptor binding domain was amplified and then cloned into the lactococcus lactis expression system pNBC1000 and pNBC2000 to form pNBC1001 and pNBC2001,the confirmed sequence was cut and ligated to pTRKH2 a shuttle vector for E.coli and Lactococcus lactis to generate pNBCL1001 and pNBCL2001.... |
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| Keywords: | clostridium difficile toxin A receptor binding domain Lactococcus lactis |
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