禽多杀性巴氏杆菌外膜蛋白H受体的初步鉴定

发布时间：2018-11-02 14:59

【摘要】：目的鉴定鸡胚成纤维(CEF)细胞膜禽多杀性巴氏杆菌外膜蛋白H(OmpH)受体。方法用膜蛋白提取试剂盒制备CEF细胞的膜蛋白,通过SDS-PAGE和配体印迹检测CEF细胞膜蛋白中的OmpH受体,采用MALDI-TOF质谱技术鉴定蛋白种类,并用配体印迹、ELISA和免疫荧光技术检测OmpH受体在不同宿主食管黏膜细胞表面的分布情况。结果在转印CEF细胞膜蛋白的NC膜上有一条明显的疑似受体条带,分子量约为49kDa。MALDI-TOF质谱鉴定结果表明该受体蛋白是ATP合成酶β亚基。在转印鸡和兔食管黏膜细胞膜蛋白的NC膜上检测到能够与OmpH结合的蛋白条带,但在牛和猪食管黏膜细胞膜蛋白中未见任何印迹条带,而OmpH与鸡和兔食管黏膜细胞膜蛋白的结合力高于猪和牛食管黏膜细胞膜蛋白。结论禽多杀性巴氏杆菌OmpH受体可能是CEF细胞膜表面的ATP合成酶β亚基。
[Abstract]:Objective to identify the H (OmpH) receptor of outer membrane protein of chicken embryo fibroblast (CEF) cell membrane of Pasteurella multocida (Pasteurella multocida). Methods the membrane proteins of CEF cells were prepared by membrane protein extraction kit. The OmpH receptors in CEF cell membrane proteins were detected by SDS-PAGE and ligand blotting. The proteins were identified by MALDI-TOF mass spectrometry and ligand blotting. ELISA and immunofluorescence techniques were used to detect the distribution of OmpH receptors on the surface of esophageal mucosal cells in different hosts. Results there was an obvious suspected receptor band on the NC membrane of CEF cell membrane protein. The molecular weight of the receptor protein was identified by 49kDa.MALDI-TOF mass spectrometry. The result showed that the receptor protein was a 尾 subunit of ATP synthase. On the NC membrane of the esophageal mucosal membrane proteins of chickens and rabbits, the protein bands that could bind to OmpH were detected, but no imprinted bands were found in the membrane proteins of bovine and pig esophagus mucosa. The binding ability of OmpH to esophageal membrane protein of chicken and rabbit was higher than that of porcine and bovine esophageal mucosal membrane protein. Conclusion the OmpH receptor of Pasteurella multocida may be a ATP synthase 尾 subunit on the surface of CEF cell membrane.
【作者单位】：伊犁师范学院生物与地理科学学院;吉首大学生物资源与环境科学学院;中国疾病预防控制中心传染病预防控制所;
【基金】：国家自然科学基金项目(No.31360613)资助~~
【分类号】：S852.61

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