TN7 TRANSPOSITION INVITRO PROCEEDS THROUGH AN EXCISED TRANSPOSON INTERMEDIATE GENERATED BY STAGGERED BREAKS IN DNA
TN7 TRANSPOSITION INVITRO PROCEEDS THROUGH AN EXCISED TRANSPOSON INTERMEDIATE GENERATED BY STAGGERED BREAKS IN DNA
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DOI：
10.1016/0092-8674(91)90388-f
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发表时间：
1991-05-31
期刊：
影响因子：
64.5
通讯作者：
CRAIG, NL
中科院分区：
文献类型：
--
作者：
BAINTON, R;GAMAS, P;CRAIG, NL
我们已经开发了一个无细胞系统，其中细菌转座子Tn7高频插入到其在大肠杆菌染色体上的首选靶点attTn7；体外转座需要ATP和Tn7编码的蛋白质。Tn7通过剪切和粘贴机制进行转座，在该机制中，通过交错的双链断裂将元件从供体DNA中切除，然后通过3‘转座子末端与5’靶端的连接将其插入attTn7。在没有任何蛋白质成分或DNA底物的情况下，既不能观察到重组中间体，也不能观察到产物。因此，我们认为Tn7转座发生在包含几种蛋白质和底物DNA的核蛋白复合体中，并且在该复合体中识别attTn7会引起Tn7末端的链切割。
We have developed a cell-free system in which the bacterial transposon Tn7 inserts at high frequency into its preferred target site in the Escherichia coli chromosome, attTn7; Tn7 transposition in vitro requires ATP and Tn7-encoded proteins. Tn7 transposes via a cut and paste mechanism in which the element is excised from the donor DNA by staggered double-strand breaks and then inserted into attTn7 by the joining of 3' transposon ends to 5' target ends. Neither recombination intermediates nor products are observed in the absence of any protein component or DNA substrate. Thus, we suggest that Tn7 transposition occurs in a nucleoprotein complex containing several proteins and the substrate DNAs and that recognition of attTn7 within this complex provokes strand cleavages at the Tn7 ends.