一个由ColE1编码的基因指导质粒的进入排斥。
A ColE1-encoded gene directs entry exclusion of the plasmid.
作者信息
Yamada Y, Yamada M, Nakazawa A
机构信息
Department of Biochemistry, Yamaguchi University School of Medicine, Japan.
出版信息
J Bacteriol. 1995 Nov;177(21):6064-8. doi: 10.1128/jb.177.21.6064-6068.1995.
Abstract
To detect entry exclusion of the ColE1 plasmid, we established an assay system that was not influenced by incompatibility of extant plasmids in the recipient cells or by the viability of the cells due to the killing action of colicin E1 protein. The assay revealed that exc1 and exc2, assigned as genes directing entry exclusion, had no exclusion activity. Instead, mbeD, which had been characterized as a gene for plasmid mobilization, directed the exclusion activity. MbeD was overexpressed and identified as a 35S-labeled protein, which was recovered in both the soluble and membrane fractions, particularly in the inner membrane fraction. An amphipathic helical structure was predicted in the N-terminal region of MbeD as well as in the corresponding homologous proteins of ColA and ColK. These proteins may bind to the inner membrane via the N-terminal amphipathic helix and function in entry exclusion.
摘要
为检测ColE1质粒的进入排斥，我们建立了一个检测系统，该系统不受受体细胞中现存质粒不相容性的影响，也不受因大肠杆菌素E1蛋白的杀伤作用导致的细胞活力的影响。该检测表明，被指定为指导进入排斥的基因exc1和exc2没有排斥活性。相反，曾被鉴定为质粒转移基因的mbeD指导排斥活性。MbeD被过度表达并被鉴定为一种35S标记的蛋白质，它在可溶性和膜组分中均有回收，尤其是在内膜组分中。在MbeD的N端区域以及ColA和ColK的相应同源蛋白中预测有两亲性螺旋结构。这些蛋白质可能通过N端两亲性螺旋与内膜结合并在进入排斥中发挥作用。
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