Libraries for genomic SELEX
Libraries for genomic SELEX
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DOI：
10.1093/nar/25.4.781
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发表时间：
1997-02-15
影响因子：
14.9
通讯作者：
Gold, L
中科院分区：
文献类型：
--
作者：
Singer, BS;Shtatland, T;Gold, L
越来越多的蛋白质被鉴定为通过在体内结合特异性核酸来调节基因表达。一种称为基因组SELEX的方法通过在生物体的基因组序列内鉴定生物体的任何蛋白质的最高亲和力位点来促进蛋白质-核酸相互作用网络的快速鉴定。与其祖先随机序列核酸的SELEX一样，基因组SELEX涉及核酸的反复结合、分配和扩增。这两种方法的不同之处在于用于基因组SELEX的核酸文库的可变区来源于生物体的基因组。我们已经使用了一种快速和简单的方法来构建大肠杆菌，酿酒酵母，和人类基因组DNA PCR文库，可以用T7 RNA聚合酶转录。我们提出的证据表明，文库包含重叠插入开始在基因组内的大多数位置，使这些文库适合基因组SELEX。
An increasing number of proteins are being identified that regulate gene expression by binding specific nucleic acids in vivo. A method termed genomic SELEX facilitates the rapid identification of networks of protein-nucleic acid interactions by identifying within the genomic sequences of an organism the highest affinity sites for any protein of the organism. As with its progenitor, SELEX of random-sequence nucleic acids, genomic SELEX involves iterative binding, partitioning, and amplification of nucleic acids. The two methods differ in that the variable region of the nucleic acid library for genomic SELEX is derived from the genome of an organism. We have used a quick and simple method to construct Escherichia coli, Saccharomyces cerevisiae, and human genomic DNA PCR libraries that can be transcribed with T7 RNA polymerase. We present evidence that the libraries contain overlapping inserts starting at most of the positions within the genome, making these libraries suitable for genomic SELEX.