牦牛源多杀性巴氏杆菌ompH基因原核表达及抗原性鉴定

摘要

[目的]克隆、表达牦牛源多杀性巴氏杆菌(Pm)外膜蛋白基因H(ompH)并鉴定其抗原性.[方法]扩增牦牛源多杀性巴氏杆菌去信号肽的ompH基因,构建原核表达载体pET28a - ompH,转化大肠杆菌BL21( DE3)并诱导表达,通过SDS-PAGE鉴定表达目的蛋白,利用Western blot检测该蛋白的抗原性.[结果]成功克隆、构建了牦牛源多杀性巴氏杆菌去信号肽的pET28a - ompH原核表达载体.诱导表达蛋白约38 kDa,Western blot检测重组蛋白具有良好的抗原性.[结论]ompH基因的成功表达为重组OmpH蛋白的血清学检测方法的建立、多克隆抗体的制备及疫苗的研制奠定了基础.%[ Objective ] The study aims to clone and express the recombinant OmpH preotein of Pasteurella multocida (Pm) from yak. [ Methods ] The deleted signal peptide OmpH fragment of Pm from yak was amplified. The prokaryotic expression plasmid of pET28a - ompH was constructed. It was transformed into BL21(DE3) and was expressed. SDS -PAGE was carried out to identify the protein, and Western blot was employed to determine the reaction of the target protein with the Pm antibody. [Result] The construction of the prokaryotic expression plasmid of pET28a - ompH was successful. The expressed protein was about 38kDa and reactive in the Western blot detection. [ Conclusion ] The successful expression of OmpH gene takes a shot at further serological detection, polyclonal antibody preparation and vaccine development.