Ellis A E, do Vale A, Bowden T J, Thompson K, Hastings T S
SOAEFD Marine Laboratory, Aberdeen, UK.
FEMS Microbiol Lett. 1997 Apr 15;149(2):157-63. doi: 10.1111/j.1574-6968.1997.tb10323.x.
Using specific immunostaining of Western blots, the in vivo expression of several putative virulence factors of Aeromonas salmonicida subsp. salmonicida was demonstrated in infected muscle tissue of Atlantic salmon and rainbow trout. Three virulent isolates of A. salmonicida were used. One isolate was chosen because in vitro it was apparently a non-producer of the 70-kDa serine protease. Infected furuncle tissue was centrifuged and samples of the pellet and supernatant probed for evidence that the components of interest were bacterial cell-associated or secreted. The A-protein was detected in pelleted furuncle material but not in the supernatant. Lipopolysaccharide, both high and low molecular mass, was present in the pellet but only high molecular mass lipopolysaccharide was detected in the furuncle supernatant. Iron-regulated outer membrane proteins were detected in the furuncle pellet. The 70-kDa serine protease was detected in the furuncle supernatant of both protease-producing strains. However, whilst the protease-deficient isolate was demonstrated to produce low levels of the 70-kDa protease when grown in vitro under iron restricted conditions, none could be detected in vivo.
通过蛋白质免疫印迹法的特异性免疫染色,在感染的大西洋鲑鱼和虹鳟鱼肌肉组织中证实了杀鲑气单胞菌杀鲑亚种几种假定毒力因子的体内表达。使用了三株杀鲑气单胞菌的强毒株。选择其中一株是因为在体外它显然不产生70 kDa丝氨酸蛋白酶。将感染的疖组织离心,对沉淀和上清液样本进行检测,以寻找感兴趣的成分与细菌细胞相关或分泌的证据。在疖沉淀物质中检测到A蛋白,但在上清液中未检测到。沉淀中存在高分子量和低分子量的脂多糖,但在疖上清液中仅检测到高分子量脂多糖。在疖沉淀中检测到铁调节外膜蛋白。在两株产生蛋白酶的菌株的疖上清液中均检测到70 kDa丝氨酸蛋白酶。然而,虽然蛋白酶缺陷型菌株在体外铁限制条件下生长时被证明产生低水平的70 kDa蛋白酶,但在体内未检测到。