Smith S C, Roddick F, Ling S, Gerraty N L, Coloe P J
Department of Applied Biology, Royal Melbourne Institute of Technology, Vic., Australia.
Vet Microbiol. 1990 Jul;24(1):29-41. doi: 10.1016/0378-1135(90)90048-z.
The protein composition of 18 clinical isolates of Treponema hyodysenteriae from pigs with swine dysentery in Australia were compared by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblot analysis. Coomassie Blue stained SDS-PAGE-profiles of whole cell and outer membrane (OM) proteins demonstrated the same gel pattern among the T. hyodysenteriae isolates, particularly the OM proteins in the molecular mass (Mr) range of 30 kDa to 40 kDa. The T. hyodysenteriae isolates were categorised into two distinct groups (A and B) based on the strain-variability in the 37 kDa OM protein. Immunoblotting of whole cell proteins after SDS-PAGE using serum from rabbits and pigs immunised with known T. hyodysenteriae serotypes revealed a number of common immunoreactive bands in all isolates. LPS typing of the T. hyodysenteriae isolates by immunoblotting with the rabbit antiserum revealed one additional serotype emphasising the LPS heterogeneity among the strains isolated from geographic locations in Australia, Great Britain and the U.S.A. Immunoblotting of the OM preparations revealed several common immunoreactive polypeptides corresponding to Mr values of 34 kDa to 30 kDa among the T. hyodysenteriae and T. innocens isolates but a distinct 39 kDa found only in the T. hyodysenteriae isolates. Trypsin proteolysis of intact. T. hyodysenteriae cells caused selective loss of these and other major abundant proteins identifying the location of the 39 kDa, 36 kDa, 34 kDa and 30 kDa proteins on the cell surface and suggesting a possible role of these proteins in the pathogenesis of swine dysentery.
采用十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE）和免疫印迹分析，比较了从澳大利亚患猪痢疾的猪身上分离出的18株猪痢疾密螺旋体临床分离株的蛋白质组成。考马斯亮蓝染色的全细胞和外膜（OM）蛋白的SDS-PAGE图谱显示，猪痢疾密螺旋体分离株之间具有相同的凝胶图谱，特别是分子量（Mr）在30 kDa至40 kDa范围内的OM蛋白。根据37 kDa OM蛋白的菌株变异性，猪痢疾密螺旋体分离株被分为两个不同的组（A组和B组）。用已知猪痢疾密螺旋体血清型免疫的兔和猪的血清对SDS-PAGE后的全细胞蛋白进行免疫印迹分析，结果显示所有分离株中都有一些共同的免疫反应条带。用兔抗血清通过免疫印迹对猪痢疾密螺旋体分离株进行脂多糖（LPS）分型，结果显示又发现了一种血清型，这突出了从澳大利亚、英国和美国地理位置分离出的菌株之间LPS的异质性。对OM制剂的免疫印迹分析显示，猪痢疾密螺旋体和无害密螺旋体分离株中有几种共同的免疫反应性多肽，其Mr值在34 kDa至30 kDa之间，但仅在猪痢疾密螺旋体分离株中发现一条明显的39 kDa条带。完整的猪痢疾密螺旋体细胞经胰蛋白酶水解后，这些和其他主要丰富蛋白会选择性丢失，从而确定了39 kDa、36 kDa、34 kDa和30 kDa蛋白在细胞表面的位置，并提示这些蛋白可能在猪痢疾的发病机制中起作用。
