Molecular analysis of human chromosome 16 cosmid clones containing NotI sites.
Molecular analysis of human chromosome 16 cosmid clones containing NotI sites.
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含有 NotI 位点的人类 16 号染色体粘粒克隆的分子分析。
DOI：
10.1007/bf00431252
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发表时间：
1992
期刊：
影响因子：
--
通讯作者：
Landes,G
中科院分区：
文献类型：
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作者：
Lerner,T;Wright,G;Leverone,B;Dackowski,W;Shook,D;Anderson,MA;Klinger,K;Callen,D;Landes,G
为了测试使用克隆的NotI位点作为物理作图标记的可行性，我们筛选了跨越人类染色体(Chr)16上NotI位点的粘粒克隆。对这些克隆的荧光原位杂交分析证实了先前报道的16p13.3上的NotI位点簇。通过将克隆的NotI位点与含有基因组DNA的EcoRI和EcoRI/NotI酶切的Southern杂交，确定了基因组DNA上克隆的NotI位点的甲基化状态。这些结果表明，我们研究的6个无性系中有4个可以作为物理作图的连锁无性系。两个克隆的NotI位点在所测试的细胞系中是不能切割的。在一个克隆中，NotI位点作为一个孤立的稀有切割限制酶位点存在，而在另一个克隆中，NotI位点似乎是与岛屿相关的。
To test the feasibility of using clonedNotI sites as markers for physical mapping, we have screened for cosmid clones spanning theNotI sites on human Chromosome (Chr) 16. Fluorescence in situ hybridization analysis of these clones confirms the previously reported cluster ofNotI sites on 16p13.3. Methylation status of the clonedNotI sites on genomic DNA was established by hybridization of the cosmids to Southern blots containingEcoRI andEcoRI/NotI digest of genomic DNA. These results indicated that four of six clones included in our study can be used as linking clones for physical mapping. Two clones haveNotI sites which are not cleavable in the cell lines tested. In one clone, theNotI site exists as an isolated rare-cutting restriction enzyme site, whereas in the other clone theNotI site appears to be island-related.