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大肠杆菌mukB基因缺失突变体的抑制基因smbA的鉴定与表征

Identification and characterization of the smbA gene, a suppressor of the mukB null mutant of Escherichia coli.

作者信息

Yamanaka K, Ogura T, Niki H, Hiraga S

机构信息

Department of Molecular Cell Biology, Kumamoto University School of Medicine, Japan.

出版信息

J Bacteriol. 1992 Dec;174(23):7517-26. doi: 10.1128/jb.174.23.7517-7526.1992.

Abstract

The mukB gene encodes a protein involved in chromosome partitioning in Escherichia coli. To study the function of this protein, we isolated from the temperature-sensitive mukB null mutant and characterized 56 suppressor mutants which could grow at 42 degrees C. Ten of the mutants also showed cold-sensitive growth at 22 degrees C. Using one of the cold-sensitive mutants as host, the wild type of the suppressor gene was cloned. The cloned suppressor gene complemented all of the 56 suppressor mutations. DNA sequencing revealed the presence of an open reading frame of 723 bp which could encode a protein of 25,953 Da. The gene product was indeed detected. The previously undiscovered gene, named smbA (suppressor of mukB), is located at 4 min on the E. coli chromosome, between the tsf and frr genes. The smbA gene is essential for cell proliferation in the range from 22 to 42 degrees C. Cells which lacked the SmbA protein ceased macromolecular synthesis. The smbA mutants are sensitive to a detergent, sodium dodecyl sulfate, and they show a novel morphological phenotype under nonpermissive conditions, suggesting a defect in specific membrane sites.

摘要

mukB基因编码一种参与大肠杆菌染色体分离的蛋白质。为了研究该蛋白质的功能,我们从温度敏感型mukB缺失突变体中分离并鉴定了56个能在42℃生长的抑制突变体。其中10个突变体在22℃也表现出冷敏感生长。以其中一个冷敏感突变体为宿主,克隆了抑制基因的野生型。克隆的抑制基因弥补了所有56个抑制突变。DNA测序显示存在一个723 bp的开放阅读框,可编码一个25,953 Da的蛋白质。确实检测到了该基因产物。这个以前未被发现的基因,命名为smbA(mukB的抑制子),位于大肠杆菌染色体上4分钟处,在tsf和frr基因之间。smbA基因在22至42℃范围内对细胞增殖至关重要。缺乏SmbA蛋白的细胞停止大分子合成。smbA突变体对去污剂十二烷基硫酸钠敏感,并且在非允许条件下表现出一种新的形态学表型,表明特定膜位点存在缺陷。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7726/207461/e94bfc4894e5/jbacter00089-0038-a.jpg

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