White W B, Whipple D L, Stabel J R, Bolin C A
Leptospirosis/Mycobacteriosis Research Unit, National Animal Disease Center, USDA, Ames, IA 50010.
Am J Vet Res. 1994 Oct;55(10):1399-405.
Protein expression profiles of 10 isolates of Mycobacterium paratuberculosis, M avium 18 (formerly M paratuberculosis 18), and 1 isolate each of M avium serotype 2, M avium serotype 8, and M bovis BCG were examined. Protein expression profiles of M paratuberculosis and M avium were similar. However, two-dimensional gel analysis of [35S]methionine-labeled cellular proteins resolved 4 proteins, with molecular mass of 28,000, 32,000, 32,000, and 42,000 daltons, which were expressed in greater amounts in M paratuberculosis than in M avium. Two proteins, with molecular mass of 43,000 and 60,000 daltons, were identified, which were expressed in greater amounts in M avium than in M paratuberculosis. Immuno (western)-blot analysis, using antiserum from 2 cows clinically infected with M paratuberculosis as the primary antibodies, suggested that the 42,000-dalton protein may be specific for M paratuberculosis. Comparison of protein expression profiles may be useful as a tool for differentiating isolates of M paratuberculosis. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of [35S]methionine-labeled extracellular proteins revealed variability among the isolates. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of [35S]methionine-labeled cellular proteins divided the M paratuberculosis isolates into 2 groups on the basis of a difference in the amount of expression of a 28,000-dalton protein. This information may be useful in epidemiologic studies.
对10株副结核分枝杆菌、鸟分枝杆菌18型（以前称为副结核分枝杆菌18型）以及1株鸟分枝杆菌血清型2、鸟分枝杆菌血清型8和卡介苗（M bovis BCG）的蛋白质表达谱进行了检测。副结核分枝杆菌和鸟分枝杆菌的蛋白质表达谱相似。然而，对[35S]甲硫氨酸标记的细胞蛋白进行二维凝胶分析时，分辨出4种蛋白质，其分子量分别为28,000、32,000、32,000和42,000道尔顿，这些蛋白质在副结核分枝杆菌中的表达量高于鸟分枝杆菌。还鉴定出两种蛋白质，分子量分别为43,000和60,000道尔顿，它们在鸟分枝杆菌中的表达量高于副结核分枝杆菌。使用来自2头临床感染副结核分枝杆菌的奶牛的抗血清作为一抗进行免疫（western）印迹分析表明，42,000道尔顿的蛋白质可能是副结核分枝杆菌特有的。比较蛋白质表达谱可能作为区分副结核分枝杆菌分离株的一种工具。对[35S]甲硫氨酸标记的细胞外蛋白进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳显示，各分离株之间存在差异。对[35S]甲硫氨酸标记的细胞蛋白进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳，根据28,000道尔顿蛋白质表达量的差异将副结核分枝杆菌分离株分为两组。这一信息可能在流行病学研究中有用。
