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与副溶血性弧菌耐热直接溶血素相关的模仿弧菌溶血素的纯化与特性分析

Purification and characterization of a hemolysin of Vibrio mimicus that relates to the thermostable direct hemolysin of Vibrio parahaemolyticus.

作者信息

Yoshida H, Honda T, Miwatani T

机构信息

Research Institute for Microbial Diseases, Osaka University, Japan.

出版信息

FEMS Microbiol Lett. 1991 Dec 1;68(3):249-53. doi: 10.1016/0378-1097(91)90364-g.

Abstract

A hemolysin (designated Vm-rTDH) from Vibrio mimicus (AQ0915-E13) was purified by ammonium sulfate fractionation and successive column chromatography with DEAE-Sephadex A-25, hydroxyapatite, Mono Q, Superose 12 and Phenyl-Superose. The Mr of the subunit was estimated to be about 22,000 by sodium dodecyl sulfate-slab gel electrophoresis. The isoelectric point of Vm-rTDH was approximately pH 4.9. The hemolytic activity of Vm-rTDH was stable upon heating at 100 degrees C for 10 min, similar to that of the thermostable direct hemolysin (Vp-TDH) of V. parahaemolyticus. Vm-rTDH also showed lytic activities similar to those of Vp-TDH. Immunological cross-reactivity between Vp-TDH and Vm-rTDH was demonstrated by the Ouchterlony double-diffusion test. Thus we conclude that V. mimicus produces a newly discovered type of hemolysin (Vm-rTDH) which is similar to Vp-TDH.

摘要

用硫酸铵分级沉淀法以及先后通过DEAE-葡聚糖A-25、羟基磷灰石、Mono Q、Superose 12和苯基-琼脂糖凝胶柱色谱法,从模仿弧菌(AQ0915-E13)中纯化出一种溶血素(命名为Vm-rTDH)。通过十二烷基硫酸钠平板凝胶电泳估计该亚基的相对分子质量约为22,000。Vm-rTDH的等电点约为pH 4.9。Vm-rTDH的溶血活性在100℃加热10分钟后仍保持稳定,这与副溶血性弧菌的耐热直接溶血素(Vp-TDH)相似。Vm-rTDH还表现出与Vp-TDH相似的溶解活性。通过双向免疫扩散试验证实了Vp-TDH和Vm-rTDH之间的免疫交叉反应性。因此,我们得出结论,模仿弧菌产生了一种新发现的与Vp-TDH相似的溶血素(Vm-rTDH)。

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