幽门螺杆菌唾液酸结合黏附素(SabA)的克隆表达  

Cloning and expressing of Helicobacter pylori sialic acid-binding adhesion(SabA)

作　　者：李冬宏[1,2] 宫雅楠[1] 肖迪[1] 周南进[2] 谢勇[3] 张建中[1] 

机构地区：[1]中国疾病预防控制中心传染病预防控制所诊断室,传染病预防控制国家重点实验室,北京102206 [2]南昌大学医学科学研究所,南昌330006 [3]南昌大学第一附属医院消化研究所,南昌330006

出　　处：《中国人兽共患病学报》2012年第3期197-200,共4页Chinese Journal of Zoonoses

基　　金：国家科技支撑计划(No.2007BAI04B02)资助

摘　　要：目的构建幽门螺杆菌(Helicobacter pylori,简称H.pylori或Hp)唾液酸结合黏附素(sialic acid-binding adhe-sion,SabA)的重组质粒,并在大肠杆菌BL21中表达,为探讨SabA的功能及相关疫苗的发展提供帮助。方法以Hp26695株基因组为模板,设计sabA基因特异性引物扩增目的基因,将PCR产物插入到pGEX-4T-1载体构建重组质粒,在大肠杆菌BL21中诱导表达。表达蛋白经飞行质谱鉴定,并用免疫印迹法评价其抗原性。结果经SDS-PAGE和飞行质谱鉴定,该表达蛋白为Hp外膜蛋白SabA,经免疫印法迹检测具有良好的抗原性。结论成功克隆了Hp的SabA,并能在大肠杆菌BL21中表达,该蛋白具有良好的抗原性。The aim was to construct a recombinant plasmid that expresses the Helicobacter pylori（H.pylori）sialic acid-binding adhesion（SabA） for the further study of vaccine and function of SabA.The primers were designed according to the sequence of the H.pylori 26695 sabA gene.PCR products were inserted into expression vector pGEX-4T-1,and transformed into E.coli BL21 strain for the expression.The recombinant protein was identified by MALDI-TOF-MS and the antigenicity was tested by Western blot.Results indicated that the recombinant protein was identified as H.pylori outer membrane protein SabA by SDS-PAGE and MALDI-TOF-MS,which showed good antigenicity in Western blot.It＇s concluded that the gene sabA of H.pylori 26695 was successfully cloned and expressed in E.coli BL21,and the protein expressed has good antigenicity.

关 键 词：幽门螺杆菌 唾液酸黏附素 基因克隆 表达 

分 类 号：R378.2[医药卫生—病原生物学]