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转座所需的IS1末端的DNA序列。

DNA sequence at the end of IS1 required for transposition.

作者信息

Gamas P, Galas D, Chandler M

出版信息

Nature. 1985;317(6036):458-60. doi: 10.1038/317458a0.

Abstract

The insertion sequence IS1 belongs to a class of bacterial transposable genetic elements that can form compound transposons in which two copies of IS1 flank an otherwise non-transposable segment of DNA. IS1 differs from other known elements of this class (such as IS10, IS50 and IS903) in several respects. It is one of the smallest known insertion elements, exhibits a relatively complex array of open reading frames, is present in the chromosomes of various Enterobacteria, in some cases in many copies, and its insertion can result in the duplication of either 8 or 9 base pairs (bp) in the target DNA. Furthermore, although, like other members of the compound class, it seems to undergo direct transposition, IS1 also promotes replicon fusion (co-integrate formation) at a relatively high frequency. Like all other elements studied to date, the integrity of the extremities of IS1 are essential for efficient transposition. We have constructed a test system to determine the minimal DNA sequences at the extremities of IS1 required for transposition. Sequential deletions of the end sequences reveal that 21-25 bp of an isolated extremity are sufficient for transposition. A specific sequence 13-23 bp from the ends, defining the edge of the minimal sequence, is implicated as an essential site. The sites, symmetrically arrayed at both ends of IS1, correspond to the apparent consensus sequence of the known binding sites for the Escherichia coli DNA-binding protein (called integration host factor or IHF) which is required for the site-specific recombination that leads to integration of bacteriophage lambda into the bacterial genome. The sites at the ends of IS1 may thus bind a host protein, such as JHF or a related protein, that is involved in regulating the transposition apparatus.

摘要

插入序列IS1属于一类细菌转座遗传元件,它能够形成复合转座子,其中两个IS1拷贝位于一段原本不可转座的DNA片段两侧。IS1在几个方面与该类其他已知元件(如IS10、IS50和IS903)不同。它是已知最小的插入元件之一,具有相对复杂的开放阅读框阵列,存在于各种肠道杆菌的染色体中,在某些情况下有多个拷贝,其插入可导致靶DNA中8个或9个碱基对(bp)的重复。此外,尽管与复合类的其他成员一样,它似乎进行直接转座,但IS1也以相对较高的频率促进复制子融合(共整合体形成)。与迄今研究的所有其他元件一样,IS1末端的完整性对于有效转座至关重要。我们构建了一个测试系统,以确定IS1末端转座所需的最小DNA序列。对末端序列进行连续缺失分析表明,一个孤立末端的21 - 25 bp足以进行转座。从末端起13 - 23 bp处的一个特定序列,定义了最小序列的边缘,被认为是一个必需位点。这些位点在IS1两端对称排列,对应于大肠杆菌DNA结合蛋白(称为整合宿主因子或IHF)已知结合位点的明显共有序列,该蛋白是噬菌体λ整合到细菌基因组中所需的位点特异性重组所必需的。因此,IS1末端的这些位点可能结合一种宿主蛋白,如IHF或相关蛋白,该蛋白参与调节转座装置。

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