伤寒沙门菌高渗诱导基因osmY的克隆表达与多克隆抗体的制备  

Gene cloning,expression and polyclonal antibody preparation of osmotically inducible gene osmY of Salmonella enterica serovar Typh

作　　者：翁晓琴[1] 张红[1] 詹莉芳[1] 张晓磊[1] 生秀梅[1] 徐顺高[1] 黄新祥[1] 

机构地区：[1]江苏大学基础医学与医学技术学院,江苏镇江212013

出　　处：《江苏大学学报（医学版）》2013年第3期243-246,共4页Journal of Jiangsu University：Medicine Edition

基　　金：国家自然科学基金资助项目(31000046);江苏大学高级人才启动基金资助项目(11JDG063)

摘　　要：目的:克隆表达伤寒沙门菌高渗诱导基因osmY并制备多克隆抗体。方法:通过PCR技术从伤寒沙门菌基因组DNA中获得伤寒沙门菌高渗诱导基因osmY,再将此基因片段克隆到表达载体pET22b(+)上,并转入大肠埃希菌JM109进行表达;Ni柱纯化后的OsmY蛋白作为抗原免疫家兔,并获得多克隆抗体。结果:成功制备伤寒沙门菌高渗诱导基因osmY的蛋白表达菌株,并获得了纯化的OsmY蛋白,成功制备兔抗OsmY的多克隆抗体。结论:成功克隆表达了伤寒沙门菌高渗诱导基因osmY,并获得了多克隆抗体,有助于今后研究OsmY在伤寒沙门菌高渗应激中的作用。Objective： To clone and express osmotically inducible gene osmY of Salmonella enterica serovar Typhi（S.Typhi）,and prepare the anti-OsmY polyclonal antibody.Methods： The DNA fragment of osmotically inducible gene osmY from S.Typhi GIFU 10007 was cloned using PCR.The sequence encoding the protein OsmY was cloned into the pET22b（＋） vector and expressed in E.coli JM109.The fusion protein OsmY-His6 was purified by Ni affinity chromatography and was used as antigen to prepare polyclonal antibody.Results： The protein expression strain of osmotically inducible gene osmY was prepared successfully and was highly expressed in E.coli JM109.The anti-OsmY polyclonal antibody was prepared successfully. Conclusion： This study was contributed to research on the role of OsmY in hyperosmotic stress of S.Typhi.

关 键 词：伤寒沙门菌 osmY 多克隆抗体 高渗诱导基因 

分 类 号：R387.22[医药卫生—医学寄生虫学]