The genetic analysis of recombination using adenovirus overlapping terminal DNA fragments.
The genetic analysis of recombination using adenovirus overlapping terminal DNA fragments.
复制标题
使用腺病毒重叠末端DNA片段进行重组的遗传分析。
DOI：
10.1016/0042-6822(83)90072-7
复制
发表时间：
1983
期刊：
影响因子：
3.7
通讯作者：
Young,CS
中科院分区：
文献类型：
--
作者：
Volkert,FC;Young,CS
我们已经研究了腺病毒基因组重叠末端片段之间的遗传重组的后果，相对于重叠序列中的标记。研究结果是一致的，一般重组发生约同位素的间隔内。特别是，单一标记内的重叠，评分非选择性，显示频率的恢复依赖于其位点的位置相对于重叠的两端。成对的ts标记重组形成ts+后代，与它们的距离成比例，条件是标记被定向，使得它们之间的单个交换将产生携带两个ts+等位基因的全长基因组。在相反的方向上，这样一个单一的交叉将被预期产生ts ts重组体，ts+频率低得多，表明多个重组事件是罕见的，在这个系统中。这些发现排除了位点特异性重组、仅在重叠的切割末端发生的重组以及通过重叠长度的异源双链体的错配修复进行的重组。它们还表明，在该反应过程中形成的任何异源双链体接合区都非常短，或者它不受异源双链体修复的影响。最后，我们的研究结果表明重叠重组作为遗传和物理作图工具和作为菌株构建方法的有效性，并且它们建议其他应用，例如使用重叠重组来证明紧密间隔的标记对（例如，假定的第二位点回复及其伴随的是病变）可以被分离。
We have studied the consequences of genetic recombination between overlapping terminal fragments of adenovirus genomes with respect to markers in the overlapping sequence. The findings are consistent with general recombination occurring approximately isotonically within the interval. In particular, single markers within the overlap, scored nonselectively, showed frequencies of recovery dependent on the position of their locus in relation to the ends of the overlap. Pairs of ts markers recombined to form ts+ progeny in proportion to their distance apart, provided the markers were oriented so that a single crossover between them would produce a full-length genome bearing both ts+ alleles. In the opposite orientation, where such a single crossover would be expected to produce ts ts recombinants, the ts+ frequency was much lower, indicating that multiple recombination events are rare in this system. These findings rule out site-specific recombination, recombination occurring exclusively at the cleaved ends of the overlap, and recombination by means of mismatch repair of a heteroduplex the length of the overlap. They also indicate either that any heteroduplex junction region formed in the course of this reaction is quite short or that it is not subject to heteroduplex repair. Finally, our results demonstrate the efficacy of overlap recombination as a genetic and physical mapping tool and as a method of strain construction, and they suggest other applications, such as using overlap recombination to demonstrate that closely spaced pairs of markers (eg, putative second-site reversions and their accompanying is lesions) can be segregated.