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首页> 外文期刊>Infection and immunity >Investigation of a hemolysin produced by enteropathogenic Treponema hyodysenteriae.
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Investigation of a hemolysin produced by enteropathogenic Treponema hyodysenteriae.

机译：肠道致病性痢疾密螺旋体产生的溶血素的研究。

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A hemolysin produced by Treponema hyodysenteriae, the etiological agent of swine dysentery, was investigated. A virulent isolate (B204) was inoculated into a standard culture medium consisting of Trypticase soy broth without dextrose (BBL Microbiology Systems) supplemented with 10% fetal calf serum in an atmosphere of 70:30 deoxygenated H2-CO2. Sterile cell-free filtrates were prepared at 2-h intervals and assayed for hemolytic activity by using washed sheep erythrocytes. The maximum hemolytic titer was obtained during the early log phase of growth (4 h). A loss of hemolytic activity was observed when cell-free filtrates were stored at 23 and 4 degrees C. Storage at -20 or -80 degrees C after lyophilization resulted in retention of the hemolytic titer for periods of up to 30 days. Enzymatic inactivation of the hemolysin was accomplished with pronase, but not with deoxyribonuclease, ribonuclease, lipase, or trypsin. Addition of exogenous ribonucleic acid-core to the standard culture medium resulted in a dose-dependent increase in the amount of hemolysin produced. The hemolysin could be purified by acid and ammonium sulfate precipitation followed by ion exchange and molecular sieve chromatography. The molecular weight of the hemolysin was 68,000 when determined by sodium dodecyl sulfate-polyacrylamide slab gel electrophoresis.
机译：对猪痢疾的病原体猪痢疾螺旋体产生的溶血素进行了研究。在70:30脱氧的H2-CO2气氛中，将强毒分离株（B204）接种到由不含葡萄糖的胰蛋白酶的大豆肉汤组成的标准培养基（BBL Microbiology Systems）中，并添加10％的小牛血清。每隔2小时制备无菌的无细胞滤液，并使用洗涤过的绵羊红血球测定溶血活性。在生长的早期对数期（4 h）获得最大溶血滴度。当将无细胞的滤液储存在23和4摄氏度时，观察到溶血活性的丧失。冻干后在-20或-80摄氏度下储存导致溶血滴度保持长达30天。溶血素的酶失活是通过链霉蛋白酶完成的，而不是通过脱氧核糖核酸酶，核糖核酸酶，脂肪酶或胰蛋白酶完成的。将外源核糖核酸核心添加到标准培养基中导致溶血素产生量的剂量依赖性增加。溶血素可以通过酸和硫酸铵沉淀，离子交换和分子筛色谱法纯化。通过十二烷基硫酸钠-聚丙烯酰胺平板凝胶电泳测定时，溶血素的分子量为68,000。

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