MECHANISM OF STRAND CLEAVAGE AND EXCHANGE IN THE CRE-LOX SITE-SPECIFIC RECOMBINATION SYSTEM
MECHANISM OF STRAND CLEAVAGE AND EXCHANGE IN THE CRE-LOX SITE-SPECIFIC RECOMBINATION SYSTEM
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DOI：
10.1016/0022-2836(85)90224-4
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发表时间：
1985-01-01
影响因子：
5.6
通讯作者：
ABREMSKI, K
中科院分区：
文献类型：
--
作者：
HOESS, RH;ABREMSKI, K
噬菌体P1重组酶Cre介导loxP位点之间的位点特异性重组。loxP位点由两个由8个碱基对间隔区分隔的13个碱基对反向重复序列组成。当含有loxP位点的DNA与Cre孵育时，在间隔区内发生特异性切割，产生6个碱基对的交错切割。切口以loxP位点的二分体对称轴为中心，产生5“突出末端：图形 **。在裂解点，Cre变成共价连接到3“PO4，并产生游离的5”OH。进行了一系列实验，其中放射性标记的loxP位点与未标记的loxP位点重组，以定位重组过程中发生链交换的点。链交换的点与检测到DNA骨架的Cre切割的位点一致。
The bacteriophage P1 recombinase Cre mediates site-specific recombination between loxP sites. The loxP site consists of two 13-base-pair inverted repeats separated by an 8-base-pair spacer region. When DNA containing the loxP site is incubated with Cre, specific cleavages occur within the spacer region, creating a 6-base-pair staggered cut. The cuts are centered on the axis of dyad symmetry of the loxP site, resulting in a 5'' protruding terminus: .**GRAPHIC**. At the point of cleavage, Cre becomes covalently attached to a 3'' PO4, and produces a free 5'' OH. A series of experiments were carried out in which a radioactively labeled loxP site is recombined with an unlabeled loxP site to locate the point at which strand exchange takes place during recombination. The points of strand exchange coincide with the sites at which Cre cleavage of the DNA backbone was detected.