禽源多杀性巴氏菌外膜蛋白的提取及其对小鼠的免疫原性观察

Extraction of Outer Membrane Protein form Pasteurella multocida and Observations on Its Antigenicity for Mice

用EDTA—溶菌酶系统处理无荚膜的5:A型禽多杀性巴氏菌,得到无粘肽层的细胞膜.蔗糖密度梯度离心,将细胞膜分成3部分.琥珀酸脱氢酶、还原型辅酶I-2,6 二氯酚靛酚(NADH-DICP)氧化还原酶和NADH氧化酶的活性以及脂多糖含量测定表明,这3部分分别是细胞内膜、细胞外膜和混合膜.用高压液相色谱法从分离的外膜中提纯外膜蛋白,获得2个主要成分P_1和P_2.将P_1和P_2分别免疫小鼠,其保护率分别为20%和90%.SDS-聚丙烯酰胺凝胶电泳表明,P_2是一种主要的外膜蛋白,分子量为52000.

The uncapsular strains of 5:A serotype of Pasteurella multocida were treated with EDTA - lysozyme system to prepare peptidoglycan-less cell membrane . After sucrose density gradient centrifugation, the cell membrane constituent was divided into three fractions , L, M and H. Assays of their succinate dehydrogenase, NADH -DICP oxido-reductase, and NADH oxidase activities, LPS content and density indicated that they are cytoplasmic, mixed and outer membrane respectively. Two major outer membrane proteins P1 and P2 were obtained by HPLC. The protective rate of P2 whose molecular weight is 52000 determined by SDS-PAGE is 90% for mice.