PURIFICATION OF CPG ISLANDS USING A METHYLATED DNA-BINDING COLUMN
PURIFICATION OF CPG ISLANDS USING A METHYLATED DNA-BINDING COLUMN
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DOI：
10.1038/ng0394-236
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发表时间：
1994-03-01
期刊：
影响因子：
30.8
通讯作者：
BIRD, AP
中科院分区：
文献类型：
--
作者：
CROSS, SH;CHARLTON, JA;BIRD, AP
CpG岛是一小段DNA，含有高密度的非甲基化CpG二核苷酸，主要与编码区相关。我们构建了一个含有大鼠染色体蛋白MeCP2的甲基-CpG结合域的亲和基质，连接到固体载体上。含有基质的柱会根据DNA的CpG甲基化程度来分离DNA，强烈保留那些高度甲基化的序列。利用这个柱子，我们开发了一种从人类基因组DNA中大量分离CpG岛的程序。由于CpG岛与大约60%的人类基因重叠，所得到的CpG岛文库可用于分离全长cDNA并将基因放置在基因组图谱上。
CpG islands are short stretches of DNA containing a high density of non-methylated CpG dinucleotides, predominantly associated with coding regions. We have constructed an affinity matrix that contains the methyl-CpG binding domain from the rat chromosomal protein MeCP2, attached to a solid support. A column containing the matrix fractionates DNA according to its degree of CpG methylation, strongly retaining those sequences that are highly methylated. Using this column, we have developed a procedure for bulk isolation of CpG islands from human genomic DNA. As CpG islands overlap with approximately 60% of human genes, the resulting CpG island library can be used to isolate full-length cDNAs and to place genes on genomic maps.