猪肺炎支原体膜蛋白的电泳分析及免疫印迹检测

以猪肺炎支原体Z株为试验材料,采用SDS-PAGE和Western印迹对该菌膜蛋白进行了分析。研究表明:猪肺炎支原体经A_(26)液体培养基培养,15000 r/min离心收集菌体细胞,低渗超声法破膜获得膜制剂后,应用12.5％的凝胶对膜蛋白进行SDS-PAGE分离,在电泳图谱上呈现出36条蛋白带,相对分子质量范围为1.18×10~4～9.12×10~4。同时以膜制剂作为抗原,分4次免疫试验兔,心脏采血,提取抗血清,并分离纯化得到IgG,然后进行Western印迹法检测。其结果发现在SDS-PAGE分离得到的36种膜蛋白(或多肽)中,有6种蛋白具有免疫原性。

Electrophoresis and Western blotting of the membrane proteins of Mycoplasma hyopneumonia

Mycoplasma hyopneumonia (Mhp) is the pathogen of swine epidemic pneumonia. It was reported that many kinds of membrane proteins of Mhp have antigenicity. In this study, Z strain of Mycoplasma hyopneumonia was incubated in liquid medium and centrifuged at 15 000 r/min to harvest the microbe. The membrane proteins of this strain were extracted by low osmosis pressure and ultrasonification and seperated by SDS-PAGE. More than 36 protein bans, with molecular weight ranged from 1. 18 ×104-9. 12×104 , were shown on the gel. However, immune blot indicated that six kinds of protein had immunogenicity.