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.
出处
《新疆农业科学》
CAS
CSCD
北大核心
2012年第1期146-149,共4页
Xinjiang Agricultural Sciences
基金
新疆农垦科学院科技引导计划(YYD201107)
新疆农垦科学院青年基金(YQJ2009-02
YQJ201110)
