Detection of Vibrio (Listonella) Anguillarum Porin Homologue Proteins and Their Source Bacteria from Coastal Seawater
Detection of Vibrio (Listonella) Anguillarum Porin Homologue Proteins and Their Source Bacteria from Coastal Seawater
复制标题
近海海水中鳗弧菌孔蛋白同源蛋白及其源菌的检测
DOI：
10.1023/a:1011109329434
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发表时间：
2000
期刊：
影响因子：
--
通讯作者：
E. Tanoue
中科院分区：
文献类型：
--
作者：
N. Yamada;S. Suzuki;E. Tanoue
本文首次报道了高知县天之内湾近海环境海水中溶解蛋白的分子分布。用抗鳗弧菌（Vibrio（Listonella）anguillarum）孔蛋白外膜蛋白（Omp 35 La）的抗体（抗Omp 35 La抗体）作探针，检测了鳗弧菌（Vibrio（Listonella）anguillarum）的细菌源溶解蛋白及其来源细菌。从沿海海水溶解的蛋白质的电泳图谱显示，大量的离散和个别的蛋白质相互重叠在一个广泛的分子量范围内，表明在沿海环境中的活性过程中，从生物体的蛋白质转移到无生命的溶解的蛋白质池。在溶解的蛋白质中，37 kDa-和18 kDa-蛋白质与Omp 35 La反应。为了分离这些溶解蛋白的来源细菌，通过用抗Omp 35 La的菌落Western印迹法筛选来自海水和病鱼的细菌。在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）/蛋白质印迹中进一步检查反应性菌株，以验证此类菌株的外膜蛋白中Omp 35 La同源物的存在。在殖民地Westernblotting阳性的129株菌中，仅4株检出与抗Omp 35 La反应的外膜蛋白。37 kDa-和18 kDa-溶解蛋白质的可能来源细菌的水平被认为比细菌总数低5-6个数量级。目前的研究使我们假设，一小部分的细菌组合是负责溶解的蛋白质在沿海沃茨。
The molecular distribution of dissolved proteins in seawater from coastal marine environments in Uranouchi Bay, Kochi Prefecture, is first reported in this article. Occurrence of bacteria-derived dissolved proteins and their source bacteria were examined using a probe of the antibody (anti-Omp35La) against a porin outer membrane protein (Omp35La) of the fish pathogenic bacteriumVibrio(Listonella)anguillarum. The electrophoretograms of dissolved proteins from coastal seawater showed a large number of discrete and individual proteins overlapped each other over a wide range of molecular masses indicating active processes in coastal environments in transferring proteins from organisms to the inanimate dissolved protein pool. Among the dissolved proteins, 37 kDa- and 18 kDa-proteins reacted with the Omp35La. In order to isolate the source bacteria of such dissolved proteins, bacteria from seawater and diseased fish were screened by colony Western blotting with anti-Omp35La. The reactive strains were further examined in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE)/Western blotting to verify the presence of Omp35La homologues among the outer membrane proteins of such strains. Outer membrane proteins reacting with anti-Omp35La were detected in only 4 strains of the 129 strains that were positive in the colony Western blotting. The level of possible source bacteria of 37 kDa- and 18 kDa-dissolved proteins was suggested to be 5–6 orders of magnitude lower than the total bacterial count. The present study leads us to hypothesize that a minor portion of the bacterial assemblage is responsible for the dissolved proteins in the coastal waters.