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大肠杆菌位点特异性DNA倒位的宿主因子:fis基因的克隆与特性分析

Escherichia coli host factor for site-specific DNA inversion: cloning and characterization of the fis gene.

作者信息

Koch C, Vandekerckhove J, Kahmann R

机构信息

Max-Planck-Institut für Molekulare Genetik, Otto-Warburg-Laboratorium, Berlin, Federal Republic of Germany.

出版信息

Proc Natl Acad Sci U S A. 1988 Jun;85(12):4237-41. doi: 10.1073/pnas.85.12.4237.

Abstract

The Escherichia coli (Es. coli) protein Fis (factor for inversion stimulation) stimulates site-specific DNA inversion of the G segment in phage Mu by binding to a recombinational enhancer. By using synthetic oligonucleotides deduced from the amino-terminal amino acid sequence, we have cloned the gene (termed fis) encoding this specific DNA-binding protein. The DNA sequence shows that the Fis protein is basic and contains 98 amino acids. A helix-turn-helix sequence motif characteristic of many DNA-binding proteins is located at the carboxyl-terminal end of the protein. By marker exchange, we have constructed an insertion mutation of fis. Fis is nonessential for Es. coli growth; however, inversion of the G segment of a Mu prophage was not detected in the fis mutant. The fis gene is located between 71 and 72 min on the Es. coli genetic map.

摘要

大肠杆菌(Es. coli)的Fis蛋白(倒位刺激因子)通过与重组增强子结合,刺激噬菌体Mu中G片段的位点特异性DNA倒位。利用从氨基末端氨基酸序列推导出来的合成寡核苷酸,我们克隆了编码这种特异性DNA结合蛋白的基因(称为fis)。DNA序列表明,Fis蛋白呈碱性,含有98个氨基酸。许多DNA结合蛋白特有的螺旋-转角-螺旋序列基序位于该蛋白的羧基末端。通过标记交换,我们构建了fis的插入突变体。Fis对大肠杆菌生长并非必需;然而,在fis突变体中未检测到Mu原噬菌体G片段的倒位。fis基因位于大肠杆菌遗传图谱上71至72分钟之间。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e37d/280402/8d79c955c773/pnas00264-0132-a.jpg

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