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A classical strain of Vibrio cholerae with diminished ability to process the proteolytically sensitive site in the A subunit of cholera toxin.

机译:霍乱弧菌的经典菌株其加工霍乱毒素A亚基中蛋白水解敏感位点的能力降低。

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摘要

Vibrio cholerae O1, No. 31, a strain isolated from a patient with mild diarrhea, produced mainly the unnicked cholera toxin. The amount of toxin that had accumulated in the cells was approximately 200 times lower than that secreted into the culture medium. When the unnicked toxin was purified by three successive column chromatographies and then extracted from the polyacrylamide gel, the unnicked toxin showed two bands corresponding to the A and B subunits by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and the A1 fragment was detected by trypsinization. Biological and enzymatic activities of the purified toxin with trypsinization were identical to those of cholera toxin from V. cholerae 569B as seen in the rabbit skin permeability test and the NAD:agmatine ADP-ribosyltransferase assay. DNA sequences of the A and B subunits were identical to those of the A- and B-subunit genes from the El Tor 2125 and classical 0395 strains, respectively. These data suggest that the wild V. cholerae strain, No. 31, produces a toxin identical to toxins previously reported in the literature and secretes it without accumulation in the cell, as is the case with other strains. However, strain No. 31's ability to nick the toxin is diminished compared with such abilities of other strains.
机译:霍乱弧菌O1(No. 31)是从轻度腹泻患者中分离出的一种菌株,主要产生无缺口的霍乱毒素。细胞中积累的毒素量比分泌到培养基中的毒素量低约200倍。当无缺口毒素通过三个连续的柱色谱法纯化后从聚丙烯酰胺凝胶中提取时,无缺口毒素通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳显示对应于A和B亚基的两个条带,并通过胰蛋白酶消化检测到A1片段。如在兔皮肤渗透性试验和NAD:胍丁胺ADP-核糖基转移酶测定中所见,经胰蛋白酶消化的纯化毒素的生物和酶活性与霍乱弧菌569B的霍乱毒素相同。 A和B亚基的DNA序列分别与El Tor 2125和经典0395菌株的A和B亚基基因相同。这些数据表明,野生霍乱弧菌菌株No.31产生的毒素与先前文献中报道的毒素相同,并且分泌该毒素而不会在细胞中积累,就像其他菌株一样。然而,与其他菌株的这种能力相比,第31号菌株产生毒素的能力降低了。

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