首页> 外文期刊>Genetics: A Periodical Record of Investigations Bearing on Heredity and Variation >Distribution of exchanges upon homologous recombination of exogenous DNA in Xenopus laevis oocytes.
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Distribution of exchanges upon homologous recombination of exogenous DNA in Xenopus laevis oocytes.

机译:爪蟾卵母细胞中外源DNA同源重组后交换的分布。

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Homologous recombination between DNA molecules injected into Xenopus oocyte nuclei was investigated by examining the recovery of information from differentially marked parental sequences. The injected recombination substrate was a linear DNA with terminal direct repeats of 1246 bp; one repeat differed from the other by eight single base-pair substitutions, distributed throughout the region of homology, each of which created or destroyed a restriction enzyme site. Recombination products were recovered and analyzed for their content of the diagnostic sites, either directly by Southern blot-hybridization or after cloning in bacteria. The majority (76%) of the cloned products appeared to be the result of simple exchanges-i.e., there was one sharp transition from sequences derived from one parent to sequences derived from the other. These simple exchanges were concentrated near the ends of the homologous interval and, thus, near the sites of the original molecular ends. Placing marked sites on only one side of the homologous overlap showed that marker recovery was governed largely by the positions of the molecular ends and not by the markers themselves. When a terminal nonhomology was present at one end of the substrate, the yield of recombinants was sharply decreased, but the pattern of exchanges was not affected, suggesting that products from end-blocked substrates arise by the same recombination pathway. Because of considerable evidence supporting a nonconservative, resection-annealing mechanism for recombination in oocytes, we interpret the distribution of exchanges as resulting from long-patch repair of extensive heteroduplex intermediates.
机译:通过检查从差异标记的亲本序列中回收的信息,研究了注入爪蟾卵母细胞核的DNA分子之间的同源重组。注入的重组底物是线性DNA,末端直接重复序列为1246 bp。一个重复与另一个重复的不同之处是分布在整个同源区域中的八个单碱基对取代,每个取代都产生或破坏了限制性酶切位点。回收重组产物并直接通过Southern印迹杂交或克隆到细菌中后分析其诊断部位的含量。大部分(76%)克隆产物似乎是简单交换的结果-即从一个亲本衍生的序列向另一个亲本衍生的序列有一个急剧的转变。这些简单的交换集中在同源间隔的末端附近,因此,在原始分子末端的位置附近。将标记的位点仅放置在同源重叠的一侧表明,标记物的恢复主要由分子末端的位置决定,而不是由标记物本身决定。当在底物的一端存在末端非同源性时,重组子的产量急剧下降,但是交换的模式没有受到影响,这表明来自末端封闭的底物的产物是通过相同的重组途径产生的。由于有大量证据支持卵母细胞重组的非保守,切除退火机制,我们将交换的分布解释为广泛异源双链体中间体的长补片修复。

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