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首页> 外文期刊>Journal of Clinical Microbiology >DNA homology and immunological cross-reactivity between Aeromonas hydrophila cytotonic toxin and cholera toxin.
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DNA homology and immunological cross-reactivity between Aeromonas hydrophila cytotonic toxin and cholera toxin.

机译：嗜水气单胞菌胞质毒素和霍乱毒素之间的DNA同源性和免疫交叉反应性。

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DNA colony hybridization with three 18- to 20-base-long synthetic oligonucleotide probes for cholera toxin (CT) was used to screen 12 clinical isolates of Aeromonas hydrophila. Under stringent hybridizing (overnight at 40 degrees C) and washing (1 h at 50 degrees C) conditions, nine strains reacted with the 32P-labeled CT probes. Concentrated (10x) cell-free supernatants or lysates from eight cultures, heated at 56 degrees C for 20 min, produced cytotonic effects in Y-1 mouse adrenal cells and Chinese hamster ovary (CHO) cells and caused a 1.5- to 22-fold increase in production of cyclic AMP in CHO cells. Preincubation with anti-CT reduced the CHO cell titer of cell lysates by 10-fold. In the GM1 ganglioside enzyme-linked immunosorbent assay, heated supernatants and lysates gave readings equivalent to 3.5 to 100 ng of CT. Three proteins with molecular weights of 89,900, 37,000, and 11,000 reacted with anti-CT on immunoblots of cell lysates from sodium dodecyl sulfate-polyacrylamide gels. These results suggest that there is DNA homology and immunological cross-reactivity between CT and the A. hydrophila cytotonic toxin.
机译：使用三个针对霍乱毒素（CT）的18至20个碱基长的合成寡核苷酸探针进行DNA菌落杂交，以筛选12种嗜水气单胞菌临床分离株。在严格杂交（在40摄氏度过夜）和洗涤（在50摄氏度1小时）条件下，九个菌株与32P标记的CT探针反应。来自八种培养物的浓缩（10x）无细胞上清液或裂解物，在56°C加热20分钟，在Y-1小鼠肾上腺细胞和中国仓鼠卵巢（CHO）细胞中产生胞吞作用，并引起1.5至22倍的折叠增加CHO细胞中环状AMP的产生。与抗CT的预孵育将细胞裂解物的CHO细胞滴度降低了10倍。在GM1神经节苷脂酶联免疫吸附试验中，加热的上清液和裂解液的读数相当于3.5至100 ng CT。三种分子量分别为89,900、37,000和11,000的蛋白质与抗CT反应，对十二烷基硫酸钠-聚丙烯酰胺凝胶细胞裂解液的免疫印迹反应。这些结果表明，CT和嗜水链球菌胞质毒素之间存在DNA同源性和免疫交叉反应性。

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