嗜肺巴氏杆菌蛋白及抗原图谱初步分析

对不同来源的 1 1株嗜肺巴氏杆菌进行了全菌可溶性蛋白及抗原图谱分析。使用SDS -聚丙烯酰胺凝胶电泳 (SDS -PAGE) ,梯度凝胶电泳结果显示 ,嗜肺巴氏杆菌蛋白主要分布于相对分子质量 ( 1 4 4～ 97 4)× 1 0 3,且带型基本一致。 6株菌与参考菌株有完全相同的蛋白带 ,另外 5株则缺乏 40× 1 0 3带。分别用嗜肺巴氏杆菌免疫血清和自然感染嗜肺巴氏杆菌小鼠血清对 1 1株菌进行了免疫印迹 (Westernblots)试验。与免疫小鼠血清的反应显示 ,1 1株受试菌主要有相对分子质量大约 ( 1 7、31 )× 1 0 3两条反应带 ;在与自然感染血清的反应中主要的反应带在 1 7× 1 0 3处 ,缺乏 40× 1 0 3蛋白的 5株菌有 31× 1 0 3反应带 ,其余 7株均有大约 ( 2 0、2 8)× 1 0 3的反应带 ,故可以认为该菌主要抗原大约为 ( 1 7、31 )× 1 0 3;1 0个流行株根据 40× 1 0 3蛋白和 ( 2 0、2 8)× 1 0 3抗原的有无可被分为两型。本研究为精制血清学方法的诊断抗原 ,以及将SDS -PAGE和Westernblot法用于嗜肺巴氏杆菌检测奠定了基础。

Primary Analysis of Protein and Antigen Mapping of Pasteurella Pneumotropica

The pr otein and antigen mapping of 11 isolates of Pasteurlla pn eumotropica from different of breed facilities were analyzed. The results of 5-2 0% and 10-20% SDS-PAGE showed that there are 25-30 protein bands between 14.4-97 .4kD. The 12% SDS-PAGE revealed the major protein bands evident at approximately 31,34 and 42.7kDa in all isolates tested. 6 of which have the same mapping comp are with the reference strain, 5 are lack of 40kDa band. Western bolt analyses i ndicated the major antigen at approximately 17 and 31kDa in all isolates tested (the reference stain included), with the antisera of P.pneumotropica. Same test with mice sera of the naturally infected by P.pneumotropica, the major antigen at approximately 17kDa. 5 isolates tested lacked of 40kDa protein showed 31kDa b ands, and the others revealed 20 and 28kDa bands. This study demonstrated that t he different of the isolates of P.pneumotropica have the similar protein banding patterns. most of them are at the approximately 31, 34 and 42.7kDa. The major antigen are 17 and 31kD protein. Two types of P.pneumotropica could be classif ied according to 40kDa protein and 20, 28kDa antigen. The study could be the bas ic for purifying the diagnostic antigen and monitoring the P.pneumotropica with SDS-PAGE and Western blot.