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大肠杆菌多粘菌素释放的热不稳定肠毒素的纯化

Purification of the polymyxin-released, heat-labile enterotoxin of Escherichia coli.

作者信息

Evans D J, Evans D G, Richardson S H, Gorbach S L

出版信息

J Infect Dis. 1976 Mar;133 Suppl:97-102. doi: 10.1093/infdis/133.supplement_1.s97.

Abstract

The heat-labile enterotoxin of Escherichia coli strain H-10407 has been purified by use of a commercially available affinity gel (Affi-Gel 202). This gel possesses a strong and highly specific affinity for the enterotoxin released from intact E. coli cells by polymyxin B. The polymyxin-release technique could be used with fermenter-size batches of E. coli cells grown in a casamino acids-yeast extract medium. With a simple (NH4)2SO4 back-extraction step prior to affinity chromatography, large batches of E. coli enterotoxin could be processed rapidly. Affi-Gel 202-purified E. coli enterotoxin produced a single precipitin band in the presence of several different antisera against crude preparations of the toxin. The same antigen produced a precipitin band in the presence of both cholera antitoxin and antiserum to choleragenoid.

摘要

利用市售亲和凝胶(Affi-Gel 202)对大肠杆菌H-10407菌株的热不稳定肠毒素进行了纯化。这种凝胶对多粘菌素B从完整大肠杆菌细胞中释放出的肠毒素具有很强且高度特异的亲和力。多粘菌素释放技术可用于处理在酪蛋白氨基酸-酵母提取物培养基中生长的发酵罐规模的大肠杆菌细胞批次。在亲和层析之前通过简单的硫酸铵反萃取步骤,可快速处理大量的大肠杆菌肠毒素。Affi-Gel 202纯化的大肠杆菌肠毒素在几种针对该毒素粗制品的不同抗血清存在下产生单一沉淀带。相同抗原在霍乱抗毒素和类霍乱原抗血清存在下均产生沉淀带。

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