Single-strand DNA intermediates in phage lambda's red recombination pathway
Single-strand DNA intermediates in phage lambda's red recombination pathway
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DOI：
10.1073/pnas.94.7.2951
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发表时间：
1997-04-01
影响因子：
11.1
通讯作者：
Stahl, FW
中科院分区：
文献类型：
--
作者：
Hill, SA;Stahl, MM;Stahl, FW
建立了一种方法来评估在lambda的Red重组途径中出现的早期中间产物。双链断裂在体内被传递到非复制的lambda染色体上。总DNA抽提物的印迹杂交分析表明：(I)长的(1.4kb碱基)单链DNA(SsDNA)中间体；(Ii)从断裂部位双向切除；(Iii)3‘极性的单链突起；以及(Iv)在缺乏lambda’s nINR功能的情况下，红色α基因产物是生产单链DNA所必需的。因此，这些单链DNA分子表现出的物理特性与它们是先前从遗传和体外生化分析中提出的Red重组途径中的早期重组中间体是一致的。
An assay was developed to assess early intermediates arising in lambda's Red recombination pathway. Double-strand breaks were delivered in vivo to nonreplicating lambda chromosomes. Analysis by blot hybridization of total DNA extracts revealed the following: (i) long (>1.4 kilobases) single-strand DNA (ssDNA) intermediates; (ii) resection proceeding bidirectionally from the break site; (iii) single-strand overhangs of 3' polarity; and (iv) in the absence of lambda's ninR functions, a requirement of the red alpha gene product for the production of ssDNA. Therefore, the physical characteristics exhibited by these ssDNA molecules are consistent with their being an early recombination intermediate in the Red recombination pathway as proposed previously from genetic and in vitro biochemical analyses.