Overabundance of rare-cutting restriction endonuclease sites in the human genome.
Overabundance of rare-cutting restriction endonuclease sites in the human genome.
复制标题
人类基因组中存在过多的稀切限制性内切酶位点。
DOI：
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发表时间：
1987
影响因子：
14.9
通讯作者：
O. J. Miller
中科院分区：
文献类型：
--
作者：
D. Smith;W. Golembieski;J. Gilbert;L. Kizyma;O. J. Miller
从含有人3号染色体作为其唯一人类组分的体细胞杂交体构建了人3号染色体特异性粘粒文库。筛选该文库以鉴定230个含有37个酶的平均插入大小的人重组体。从54个代表2000种人类DNA内切酶的这些cosmetics中制备的DNA，然后检测EcoRI、HindIII、KpnI、XhoI和DraI的限制性内切酶位点，以及稀有切割限制性内切酶NotI、SfiI、NruI、MluI、SacII和BssHII的限制性内切酶位点。后者的酶的网站是更丰富的比预期从理论计算，反映这些网站的非随机聚类。这对使用这些酶构建染色体物理图谱具有重要意义。一些个别的染色体含有大量的稀有位点，提供了一种替代的方法，物理定位染色体的基础上确定集群的稀有限制性位点。这些簇似乎平均间隔1000 kb。
A human chromosome 3-specific cosmid library was constructed from a somatic cell hybrid containing human chromosome 3 as its only human component. This library was screened to identify 230 human recombinants which contained an average insert size of 37 kilobases. DNA prepared from 54 of these cosmids, representing 2000 kilobases of human DNA, was then tested for restriction endonuclease sites for EcoRI, HindIII, KpnI, XhoI, and DraI, as well as those of the rare-cutting restriction endonucleases NotI, SfiI, NruI, MluI, SacII, and BssHII. Sites for the latter enzymes were much more abundant than would be expected from theoretical calculations, reflecting non-random clustering of these sites. This has important implications for the use of these enzymes in the construction of physical maps of chromosomes. Some individual cosmids contained large numbers of rare sites, offering an alternative means of physically mapping chromosomes based upon identifying clusters of rare restriction sites. These clusters appear to be spaced an average of 1000 kb apart.