嗜肺巴氏杆菌外膜蛋白和脂多糖提取及其活性的初步研究  

Preliminary Study on the Extraction and Activity of Outer Membrane Protein and Lipopolysaccharide of Pasteuralla pneumotropica

作　　者：张丽芳[1] 刘星[1] 李红[1] 

机构地区：[1]中国医学科学院中国协和医科大学实验动物研究所,北京100021

出　　处：《中国比较医学杂志》2003年第5期300-304,共5页Chinese Journal of Comparative Medicine

基　　金：北京市科委实验动物科学专项基金资助( 95 3 9912 0 0 )

摘　　要：目的　提取嗜肺巴氏杆菌外膜蛋白 (OMP)和脂多糖 (LPS) ,测定其稳定性和抗原性 ,探讨其作为诊断抗原的可能性。方法　选取具有代表性的嗜肺巴氏杆菌 15株及参考株共 16株 ,提取OMP、LPS ,通过SDS -PAGE、Westernblot、ELISA等方法在稳定性、抗原性等方面与全菌抗原 (WC)进行比较。结果　 16株嗜肺巴氏杆菌OMP的SDS-PAGE图谱基本一致 ,不同培养基成分、培养时间和不同体外传代次数的提取物均显示 5条带 ,相对分子质量大约是 (17、2 6、31、37、4 1)× 10 3。其中 5株嗜肺巴氏杆菌的LPS为光滑型和粗糙型LPS的混合物 ,主带相对分子质量在 14 .4× 10 3左右。外膜蛋白和脂寡糖提取物保持了其抗原反应性和免疫原性。OMP和LPS具有种和型的特异性。OMP的 5条条带抗原性存在差异 ,相对分子质量 (2 6、37)× 10 3的蛋白不具有抗原性 ,相对分子质量 17× 10 3的蛋白抗原为受试菌株的共同抗原 ,(31、4 1)× 10 3抗原具有大鼠株和小鼠株的特异性。LPS表现出较强的型的特异性。结论　本研究提取的OMP和LPS活性稳定 ,并保持了抗原性和免疫原性。Objective By extracting Outer membrane protein(OMP) and Lipopolysaccharide(LPS) of Pasteuralla pneumotropica to study their antigenicity and stability.Methods OMP and LPS of 16 P.pneumotropica strains(including 15 representative and one reference strain) were extracted, with their antigenicity and stability compared with whole cell(WC) via ELISA, SDS-PAGE and Western Blot methods. Results OMP extracted from 16 strains of P.pneumotropica share the same result of SDS-PAGE. There're five protein bands locating at molecular weight (17, 26, 31, 37, 41)×103, which are not accessible to changes of medium component, culture time and generation. The extracted LPS of five P.pneumotropica strains are all the rough and smooth mixture,whose major band locates at molecule weight 14×103 approximately, and both antigenicity and immunity of OMP and LPS maintain. OMP and LPS have species and type specificity. There're differences between the antigenicity of five protein bands of OMP and the band at molecular weight (26, 37)×103 doesn't show antigenicity. The common antigen of five strains locates at molecular weight 17×103, and the proteins which locate at molecular weight (31, 41)×103 have different specificity between mouse strain and rat strain. LPS shows strong type specificity. Conclusion The extraction of OMP and LPS in this study are stable in activity and the antigenicity and immunity maintain.

关 键 词：嗜肺巴氏杆菌 外膜蛋白 脂多糖 提取 稳定性 抗原性 测定 

分 类 号：R37[医药卫生—病原生物学]