幽门螺杆菌外膜蛋白Omp18表达载体构建及其多克隆抗体的制备

Construction of a vector to express Helicobacter pylori outer membrane protein Omp18 and preparation of polyclonal antibodies

目的制备幽门螺杆菌的外膜蛋白Omp18及其多克隆抗体。方法设计Omp18基因引物,PCR扩增Omp18基因,将其连入pMD-18T载体,再克隆入原核表达载体pTriEx(TM)-4,重组质粒转化大肠埃希菌JM109DE,IPTG诱导表达Omp18蛋白,纯化后免疫家兔,以获得多克隆抗体。结果 PCR扩增的Omp18基因成功克隆入原核表达载体,重组原核表达载体转化菌表达Omp18蛋白,纯化的Omp18蛋白免疫动物后血清中获得多克隆抗体。结论本实验成功进行了Omp18蛋白的原核细胞表达并制备了多克隆抗体,为研究Omp18的功能、表达调控以及幽门螺杆菌检测和疫苗生产等奠定了基础。

Objective To prepare Helicobacter pylori Ompl8 protein and polyclonal antibodies against that protein Methods The Ompl8 gene was amplified using PCR and cloned into the vector pMD-18T. It was then was cloned into the prokaryotic expression plasmid vector pTriEx （TM）-4. The vector was transformed into Escherichia coli JM109DE. Ompl8 protein was produced and purified, and then rabbits were repeatedly administered the protein to produce poly- clonal antibodies. Results The Ompl8 gene was successfully cloned into the prokaryotic expression vector pTriEx （TM）-4 and the vector was successfully transformed into E. coli JM109DE. Ompl8 protein was successfully produced, and purified Omp18 protein was administered to rabbits to yield polyelonal antibodies. Conclusion Ompl8 protein was successfully expressed in prokaryotie cells, and polyclonal antibodies were successfully prepared in rabbits. This research will benefit study of the function and regulation of Ompl8 and it will lay a foundation for detection of H. pylori and prep- aration of vaccines.