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一种从赖特密螺旋体中分离鞭毛的简单方法。

A simple method for the isolation of flagella from Treponema Reiter.

作者信息

Petersen C S, Pedersen N S, Axelsen N H

出版信息

Acta Pathol Microbiol Scand C. 1981 Dec;89(6):379-85. doi: 10.1111/j.1699-0463.1981.tb02716.x.

Abstract

Flagella from Treponema Reiter were purified from a sonicate of the bacteria by diethylaminoethyl-cellulose chromatography followed by gel filtration on Sepharose CL-2B. The yield (0.8 mg flagellar protein per 8 g wet weight of treponemes) was identical to that obtained by previously described more time consuming methods. Crossed immunoelectrophoresis of the chromatographically isolated flagella against rabbit anti-T. Reiter immunoglobulin, monospecific anti-T. Reiter flagellar antibodies or human syphilitic serum in the upper gel showed two parallel but closely apposed precipitates. No contaminating material was found by electron microscopy of suspensions of flagella isolated by the new method. Sodium dodecyl sulfate polyacrylamide gel electrophoresis of dissociated flagella showed three major and one minor band. It is concluded that the new method is quantitatively and qualitatively equal to earlier described purification procedures, but simpler to use.

摘要

通过二乙氨基乙基纤维素色谱法从梅毒密螺旋体赖特尔亚种(Treponema Reiter)的超声破碎物中纯化鞭毛,随后在琼脂糖凝胶CL-2B上进行凝胶过滤。产量(每8克湿重梅毒螺旋体产生0.8毫克鞭毛蛋白)与先前描述的更耗时的方法所获得的产量相同。将经色谱分离的鞭毛与兔抗梅毒密螺旋体赖特尔亚种免疫球蛋白、单特异性抗梅毒密螺旋体赖特尔亚种鞭毛抗体或人梅毒血清在上层凝胶中进行交叉免疫电泳,显示出两条平行但紧密相邻的沉淀带。通过电子显微镜观察新方法分离的鞭毛悬浮液,未发现污染物质。解离后的鞭毛进行十二烷基硫酸钠聚丙烯酰胺凝胶电泳显示有三条主要条带和一条次要条带。得出的结论是,新方法在定量和定性方面与早期描述的纯化程序相当,但使用起来更简单。

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