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rcsF的鉴定、克隆及特性分析,rcsF是一种用于胞外多糖合成的新型调节基因,可抑制大肠杆菌K-12中的分裂突变ftsZ84。

Identification, cloning, and characterization of rcsF, a new regulator gene for exopolysaccharide synthesis that suppresses the division mutation ftsZ84 in Escherichia coli K-12.

作者信息

Gervais F G, Drapeau G R

机构信息

Department of Microbiology, Université de Montréal, Québec, Canada.

出版信息

J Bacteriol. 1992 Dec;174(24):8016-22. doi: 10.1128/jb.174.24.8016-8022.1992.

Abstract

A new gene, designated rcsF, was located adjacent to drpA at the 5.2-min position of the genetic map of Escherichia coli. The deduced amino acid sequence encoded by the rcsF gene indicates a small protein of 133 amino acid residues with a calculated pI of 10.8 that is rich in proline, serine, alanine, and cysteine residues. When overexpressed as a result of its presence on a multicopy plasmid, rcsF confers a mucoid phenotype and restores colony formation to ftsZ84 mutant cells on L agar medium containing no added NaCl. These two phenotypes are not observed in rcsB mutant cells. Ion mutant cells harboring an rcsF mutation accumulate considerably lower levels of exopolysaccharides, whereas the presence of a multicopy rcsF plasmid not only increases capsule synthesis but also confers a mucoid phenotype at 37 degrees C, a temperature at which ion mutant cells are known not to form mucoid colonies. RcsF does not stimulate the expression of rcsB, indicating that it exerts its action through the RcsB protein, possibly by phosphorylation. It is also shown that RcsF stimulation of capsule synthesis is RcsA-dependent, whereas colony formation of ftsZ84 mutant cells can be restored by RcsF in the absence of RcsA.

摘要

一个名为rcsF的新基因位于大肠杆菌遗传图谱5.2分钟位置的drpA附近。rcsF基因推导的氨基酸序列表明它是一个由133个氨基酸残基组成的小蛋白,计算所得的pI为10.8,富含脯氨酸、丝氨酸、丙氨酸和半胱氨酸残基。当rcsF因存在于多拷贝质粒上而过度表达时,它赋予细胞黏液样表型,并使ftsZ84突变细胞在未添加氯化钠的L琼脂培养基上恢复菌落形成。在rcsB突变细胞中未观察到这两种表型。携带rcsF突变的离子突变细胞积累的胞外多糖水平显著降低,而多拷贝rcsF质粒的存在不仅增加荚膜合成,还在37℃赋予黏液样表型,已知离子突变细胞在该温度下不会形成黏液样菌落。RcsF不刺激rcsB的表达,表明它可能通过磷酸化作用,经由RcsB蛋白发挥其作用。研究还表明,RcsF对荚膜合成的刺激作用依赖于RcsA,而在没有RcsA的情况下,RcsF可以恢复ftsZ84突变细胞的菌落形成。

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