METHOD OF ARTIFICIAL DNA SPLICING BY DIRECTED LIGATION (SDL)
METHOD OF ARTIFICIAL DNA SPLICING BY DIRECTED LIGATION (SDL)
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DOI：
10.1093/nar/19.24.6757
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发表时间：
1991-12-25
影响因子：
14.9
通讯作者：
BERLIN, YA
中科院分区：
文献类型：
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作者：
LEBEDENKO, EN;BIRIKH, KR;BERLIN, YA
已经设计了一种定向遗传重组的方法，它允许以预定的方式将一系列DNA片段结合在一起，以提供精确的剪接的多核苷酸序列（通过有向连接的DNA剪接，SDL）。该方法通过几种聚合酶链反应（PCR）（PCR）进行了放大，该方法利用了一组DNA片段。放大的引物包含IIS类限制性核酸内切酶的识别位点，这些核酸内切酶的识别位点将放大产物的钝端转化为独特的主要结构的突出末端，这些末端用于将片段连接在一起，是相互互补的。片段的混合物的连接，因此以明确的方式得出所需的序列。所建议的方法已通过综合编码人类成熟白介素-1-α的完全处理的（无固有的）基因来体现。
An approach to directed genetic recombination in vitro has been devised, which allows for joining together, in a predetermined way, a series of DNA segments to give a precisely spliced polynucleotide sequence (DNA splicing by directed ligation, SDL). The approach makes use of amplification, by means of several polymerase chain reactions (PCR), of a chosen set of DNA segments. Primers for the amplifications contain recognition sites of the class IIS restriction endonucleases, which transform blunt ends of the amplification products into protruding ends of unique primarY structures, the ends to be used for joining segments together being mutually complementary. Ligation of the mixture of the segments so synthesized gives the desired sequence in an unambiguous way. The suggested approach has been exemplified by the synthesis of a totally processed (intronless) gene encoding human mature interleukin-1-alpha.