A GENOMIC SCANNING METHOD FOR HIGHER ORGANISMS USING RESTRICTION SITES AS LANDMARKS
A GENOMIC SCANNING METHOD FOR HIGHER ORGANISMS USING RESTRICTION SITES AS LANDMARKS
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DOI：
10.1073/pnas.88.21.9523
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发表时间：
1991-11-01
影响因子：
11.1
通讯作者：
MUKAI, T
中科院分区：
文献类型：
--
作者：
HATADA, I;HAYASHIZAKI, Y;MUKAI, T
我们开发了一种强大的基因组扫描方法，称为“限制性标志基因组扫描”，可用于使用限制性位点作为标志来分析高等生物的基因组 DNA。基因组 DNA 在罕见切割限制性酶特异的切割位点进行放射性标记，然后在一维进行大小分级。分级分离的 DNA 进一步用另一种更常见的酶消化，并在二维中分离。该过程给出了一个二维图案，其中有数千个分散的点，对应于第一种酶的位点，表明可以以几乎等于 1 兆碱基的间隔扫描哺乳动物的基因组。基于末端标记系统的性质，斑点的位置和强度分别反映其位点和相应限制性位点的拷贝数。因此，该方法广泛适用于基因组作图或基因组改变的检测。
We have developed a powerful genomic scanning method, termed "restriction landmark genomic scanning," that is useful for analysis of the genomic DNA of higher organisms using restriction sites as landmarks. Genomic DNA is radioactively labeled at cleavage sites specific for a rare cleaving restriction enzyme and then size-fractionated in one dimension. The fractionated DNA is further digested with another more frequently occurring enzyme and separated in the second dimension. This procedure gives a two-dimensional pattern with thousands of scattered spots corresponding to sites for the first enzyme, indicating that the genome of mammals can be scanned at almost-equal-to 1-megabase intervals. The position and intensity of a spot reflect its locus and the copy number of the corresponding restriction site, respectively, based on the nature of the end-labeling system. Therefore, this method is widely applicable to genome mapping or detection of alterations in a genome.