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

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.