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编码Hin重组增强子结合蛋白的基因的分离

Isolation of the gene encoding the Hin recombinational enhancer binding protein.

作者信息

Johnson R C, Ball C A, Pfeffer D, Simon M I

机构信息

Department of Biological Chemistry, School of Medicine, University of California, Los Angeles 90024.

出版信息

Proc Natl Acad Sci U S A. 1988 May;85(10):3484-8. doi: 10.1073/pnas.85.10.3484.

Abstract

In vitro DNA inversion mediated by the protein Hin requires the presence of a recombinational enhancer sequence located in cis relative to the recombination sites and a protein, Fis, which binds to the enhancer. We have cloned and determined the primary sequence of the gene encoding Fis. The deduced amino acid sequence of Fis indicates that the protein is 98 amino acids long and contains a potential helix-turn-helix DNA binding motif at its carboxyl terminus. The gene encoding Fis maps at 72 min on the Escherichia coli chromosome. The construction of mutant strains of E. coli that lack a functional fis gene demonstrates that Fis is not essential for cell growth under laboratory conditions but is required for high rates of Hin-mediated site-specific inversion in vivo.

摘要

由蛋白质Hin介导的体外DNA倒位需要存在一个相对于重组位点顺式定位的重组增强子序列以及一种与该增强子结合的蛋白质Fis。我们已经克隆并确定了编码Fis的基因的一级序列。Fis推导的氨基酸序列表明该蛋白质由98个氨基酸组成,并且在其羧基末端含有一个潜在的螺旋-转角-螺旋DNA结合基序。编码Fis的基因定位于大肠杆菌染色体的72分钟处。构建缺乏功能性fis基因的大肠杆菌突变株表明,Fis在实验室条件下对于细胞生长不是必需的,但在体内高水平的Hin介导的位点特异性倒位中是必需的。

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