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首页> 外文期刊>Molecular and Cellular Biology >Efficient homologous recombination of linear DNA substrates after injection into Xenopus laevis oocytes.
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Efficient homologous recombination of linear DNA substrates after injection into Xenopus laevis oocytes.

机译:注射到非洲爪蟾卵母细胞中后,线性DNA底物的有效同源重组。

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When DNA molecules are injected into Xenopus oocyte nuclei, they can recombine with each other. With bacteriophage lambda DNAs, it was shown that this recombination is stimulated greatly by introduction of double-strand breaks into the substrates and is dependent on homologous overlaps in the recombination interval. With plasmid DNAs it was shown that little or no recombination occurs between circular molecules but both intra- and intermolecular events take place very efficiently with linear molecules. As with the lambda substrates, homology was required to support recombination; no simple joining of ends was observed. Blockage of DNA ends with nonhomologous sequences interfered with recombination, indicating that ends are used directly to initiate homologous interactions. These observations are combined to evaluate possible models of recombination in the oocytes. Because each oocyte is capable of recombining nanogram quantities of linear DNA, this system offers exceptional opportunities for detailed molecular analysis of the recombination process in a higher organism.
机译:将DNA分子注射到非洲爪蟾卵母细胞核中后,它们可以彼此重组。对于噬菌体λDNA,已表明,通过将双链断裂引入底物可极大地刺激这种重组,并且取决于重组间隔中的同源重叠。对于质粒DNA,显示出在环状分子之间很少或没有重组发生,但是线性分子的分子内和分子间事件都非常有效地发生。与lambda底物一样,需要同源性来支持重组。没有观察到末端的简单连接。非同源序列对DNA末端的封闭会干扰重组,这表明末端直接用于引发同源相互作用。这些观察结果相结合,以评估卵母细胞中重组的可能模型。因为每个卵母细胞都能够重组纳克级的线性DNA,所以该系统为高级生物中重组过程的详细分子分析提供了特殊的机会。

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