幽门螺杆菌ureB基因表达产物的反应原性.doc

幽门螺杆菌ureB基因表达产物的反应原性
作者:秦立篷,李丁,张菊,韩锋产,阎小君

【关键词】幽门螺杆菌
关键词: 幽门螺杆菌;尿素酶B基因;大肠杆菌
摘要:目的克隆幽门螺杆菌ureB基因,并进行表达,验证表达产物的反应原性. 方法应用PCR技术从临床分离株中扩增出ureB基因,克隆入载体pGEM-7zf(+)进行测序,进一步转到大肠杆菌表达载体pBV220进行诱导表达,以Western blot实验验证重组蛋白的反应原性. 结果测序后经同源性比较,证实扩增后得到的片断确为ureB基因,并原核表达出可被抗Hp抗体识别的非融合重组蛋白. 结论获得了有反应原性的重组UreB蛋白.

Keywords:Helicobacter pylori;urease B gene;

Abstract:AIM To clone and to express the ureB gene of Helicobacter pylori and analyse the reactionogenicity of the binant ureB fragments were amplified by PCR,then the DH5?was transformed by pBV220/ureB after sequencing and reactiono-genicity of the binant UreB was detected by Western- ureB fragments were subse-quently cloned into the prokaryotic expression vector pBV220after sequencing(The result showed that96%of the DNA se-quences we had got were the same as that of M60398,so they were the right gene we wanted).Then the DH5?was trans-fo
rmed by pBV220/ureB,and the binant protein was induced by heat when temperature reached42℃.The molec-ular mass of the expressed UreB were conformed to that we form of the binant UreB were inclu-sion bodies in host -blot showed that the binant UreB was expressed in DH5α/pBV220/ureB which could be recognized by anti- Fragment of ureB is obtained,and it is ex-pressed in