Yeast-based recombineering of DNA fragments into plant transformation vectors by one-step transformation
Yeast-based recombineering of DNA fragments into plant transformation vectors by one-step transformation
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DOI：
10.1007/s00299-007-0428-2
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发表时间：
2007-12-01
影响因子：
6.2
通讯作者：
Anai, Toyoaki
中科院分区：
文献类型：
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作者：
Nagano, Yukio;Takao, Syoko;Anai, Toyoaki
T-DNA双元载体常用于植物转化实验。因为它们通常非常大并且具有很少的适合于DNA连接反应的限制性位点，所以将DNA片段克隆到这些载体中是困难的。我们在此提供了将DNA片段克隆到非常大的载体中的替代方案。我们的基于酵母的重组工程方法使DNA片段能够通过一步转化克隆到某些类型的T-DNA二元载体中，而不需要特定的重组位点或精确定位的限制性末端，从而使克隆过程更加灵活。此外，这种方法是廉价的，适用于多片段克隆。
T-DNA binary vectors are often used in plant transformation experiments. Because they are usually very large and have few restriction sites suitable for DNA ligation reactions, cloning DNA fragments into these vectors is difficult. We provide herein an alternative to cloning DNA fragments into very large vectors. Our yeast-based recombineering method enables DNA fragments to be cloned into certain types of T-DNA binary vectors by one-step transformation without the requirement of specific recombination sites or precisely positioned restriction ends, thus making the cloning process more flexible. Moreover, this method is inexpensive and is applicable to multifragment cloning.