Recombination between linear double-stranded DNA substrates in vivo.
Recombination between linear double-stranded DNA substrates in vivo.
复制标题
线性双链 DNA 底物之间的体内重组。
DOI：
10.1016/j.ab.2009.01.015
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发表时间：
2009
影响因子：
2.9
通讯作者：
Lee,Choon-Weng
中科院分区：
文献类型：
--
作者：
Narayanan,Kumaran;Sim,EdmundUi-Hang;Ravin,NikolaiV;Lee,Choon-Weng
重组技术在大肠杆菌使线性供体DNA靶向圆形受体DNA使用短共享同源序列。在这项工作中，我们证明重组也能够在大肠杆菌体内支持一对线性DNA底物之间的重组（线性/线性重组）。高达100kb的线性DNA被精确地修饰，并且在重组后不进行重排而保持完整。该系统对于N15和PY54噬菌体和线性动物病毒等大型线性DNA的直接体内操作，以及作为人工染色体载体的线性构建体的组装具有重要价值。
Recombineering technology in Escherichia coli enables targeting of linear donor DNA to circular recipient DNA using short shared homology sequences. In this work, we demonstrate that recombineering is also able to support recombination between a pair of linear DNA substrates (linear/linear recombineering) in vivo in E. coli. Linear DNA up to 100kb is accurately modified and remains intact without undergoing rearrangements after recombination. This system will be valuable for direct in vivo manipulation of large linear DNA including the N15 and PY54 prophages and linear animal viruses, and for assembly of linear constructs as artificial chromosome vectors.