Minimum length of sequence homology required for in vivo cloning by homologous recombination in yeast
Minimum length of sequence homology required for in vivo cloning by homologous recombination in yeast
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DOI：
10.1006/plas.1997.1305
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发表时间：
1997-01-01
期刊：
影响因子：
2.6
通讯作者：
Luo, Y
中科院分区：
文献类型：
--
作者：
Hua, SB;Qiu, MS;Luo, Y
随着酿酒酵母中高效的同源重组，一种快速的体内克隆技术已经成为可能。在这里，我们证明了在DNA片段的每个末端的30 bp的同源序列足以将该片段整合到酵母中的线性化质粒中。然而，为了获得高产率的重组转化体，需要大于60 bp。有趣的是，我们观察到，在DNA片段的一端20 bp的同源性是足够的远有效的重组，只要另一端含有50 bp的同源序列。一些应用，包括高通量转移EST插入到酵母表达系统的人类基因组计划，进行了讨论。(C)北京：科学出版社.
With efficient homologous recombination in Saccharomyces cerevisiae, a rapid in vivo cloning technique has been available. Here we demonstrated that 30 bp of a homologous sequence at each end of a DNA fragment is sufficient to integrate the fragment into a linearized plasmid in yeast. To obtain a high yield of recombination transformants, however, more than 60 bp are desirable. Interestingly, we observed that 20 bp of homology at one end of a DNA fragment is sufficient far efficient recombination provided that the other end contains SO bp of homologous sequence, Some applications, including high-throughput transferring of EST inserts to the yeast expression systems for the Human Genome Project, are discussed. (C) 1997 Academic Press.