Specificity of insertion of IS1.
Specificity of insertion of IS1.
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IS1 插入的特异性。
DOI：
10.1016/0022-2836(85)90068-3
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发表时间：
1985
影响因子：
5.6
通讯作者：
Galas,D
中科院分区：
文献类型：
--
作者：
Zerbib,D;Gamas,P;Chandler,M;Prentki,P;Bass,S;Galas,D
为了确定影响靶点选择的DNA序列的特征，系统地研究了转座元件IS1插入到小的已定义序列质粒(pBR322及其衍生物)中的特异性。我们已经物理定位了几个独立的IS1插入到这些质粒中，并确定：(1)大约80%的插入发生在pBR322的唯一EcoRI位点和β-内酰胺酶基因起始之间的DNA片段(约200bp)，这是该质粒中A+T密度高的两个区域之一；(2)在这个区域有很强的定向效应(几乎所有的IS1插入都在一个方向上)，这取决于pBR322序列和供体分子中转座子的环境；(3)定向效应不依赖于pBR322中通过该区域定向的强转录。此外，我们发现将聚(da·dt)片段插入到pBR322中为IS1插入创造了一个人工热点，尽管它不像上述主要热点那样具有插入吸引力。我们的观察表明，靶序列的几个属性和供体转座子的序列环境之间的相互作用是观察到的位置和方向的特异性的原因。这里讨论的一种可能性是，转座复合体的首选“进入位点”或“信号”序列在决定IS1插入的位置和方向方面起着重要作用。
A systematic study of the specificity of insertion of the transposable element IS1 into small defined-sequence plasmids (pBR322 and derivatives) was conducted to determine the features of the DNA sequence that influence target site selection. We have physically mapped several collections of independent insertions of IS1 into these plasmids and have determined: (1) that about 80% of all insertions occur in the DNA segment (about 200 base-pairs) between the uniqueEcoRI site of pBR322 and the beginning of the β-lactamase gene, one of the two regions of high A + T density in this plasmid; (2) that there is a strong orientation effect in this region (almost all IS1 insertions are in one orientation) that depends on both the pBR322 sequence and the environment of the transposon in the donor molecule; and (3) that the orientation effect does not depend on the strong transcription that is directed through this region in pBR322. Furthermore, we have found that insertion of a poly(dA · dT) segment into pBR322 creates an artificial hotspot for IS1 insertion, even though it is not as attractive for insertion as the above-mentioned major hotspot. Our observations suggest that an interplay between several properties of the target sequences and the sequence environment of the donor transposon is responsible for the observed specificity of position and orientation. One of the possibilities discussed here is that preferred “entry-sites”, or “signal” sequences, for the transposition complex play a major role in determining the positions and orientations of IS1 insertions.
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影响因子：
5.6
作者：
GALAS, DJ;CHANDLER, M
通讯作者：
CHANDLER, M
IS1 介导的质粒 pBR322 的串联复制。
影响因子：
5.6
作者：
Chandler,M;Galas,DJ
通讯作者：
Galas,DJ
鉴定用质粒 pBR322 DNA 构建的分子嵌合体中的易位元件 IS1，其中通过聚（dA）.聚（dT）接头方法将噬菌体 MS2 DNA 拷贝插入其中。
影响因子：
5.6
作者：
R. Devos;R. Contreras;J. van Emmelo;W. Fiers
通讯作者：
W. Fiers
影响因子：
5.6
作者：
D. Galas;M. Calos;J. Miller
通讯作者：
D. Galas;M. Calos;J. Miller
通过侧翼区域的转录通读抑制插入元件 IS1 的共整合能力
影响因子：
64.5
作者：
C. Machida;Y. Machida;Hwa;K. Ishizaki;E. Ohtsubo
通讯作者：
E. Ohtsubo