转座子Tn5的靶标特异性：偏好插入G/C碱基对处。
Transposon Tn5 target specificity: preference for insertion at G/C pairs.
作者信息
Lodge J K, Weston-Hafer K, Berg D E
机构信息
Department of Microbiology and Immunology, Washington University Medical School, St. Louis, Missouri 63110.
出版信息
Genetics. 1988 Nov;120(3):645-50. doi: 10.1093/genetics/120.3.645.
Abstract
The procaryotic transposon Tn5 inserts into many different sites within a single gene, but some sites (hotspots) are targeted repeatedly. Hotspots are not closely related in sequence, but most have G/C pairs at the ends of the nine base pairs duplicated by Tn5 insertion. In pBR322, the major hotspot coincides with the "-10 region" of the tet promoter. We mutated the G/C pairs at this hotspot and assayed for insertion into hotspot I, resistance to tetracycline, and plasmid supercoiling. We found that changing the G/C pairs to A/T pairs reduced the frequency of insertion into the hotspot by at least fivefold. The reduction in hotspot use caused by these G/C to A/T changes was not attributable to changes in plasmid supercoiling or tet promoter strength.
摘要
原核转座子Tn5可插入单个基因内的许多不同位点，但有些位点（热点）会被反复靶向。热点在序列上并无紧密关联，但大多数在因Tn5插入而复制的9个碱基对末端具有G/C对。在pBR322中，主要热点与tet启动子的“-10区域”重合。我们对该热点处的G/C对进行了突变，并检测其插入热点I的情况、对四环素的抗性以及质粒超螺旋情况。我们发现，将G/C对替换为A/T对可使插入热点的频率至少降低五倍。这些由G/C到A/T的变化导致的热点使用频率降低并非归因于质粒超螺旋或tet启动子强度的变化。
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