Isolation and regional mapping of DNA sequences unique to human chromosome 21.
Isolation and regional mapping of DNA sequences unique to human chromosome 21.
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DOI：
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发表时间：
1987-12
影响因子：
9.8
通讯作者：
J. Korenberg;M. Croyle;D. Cox
中科院分区：
文献类型：
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作者：
J. Korenberg;M. Croyle;D. Cox
为了分离21号染色体特有的DNA序列，我们使用了重组DNA文库，该文库由含有21号染色体作为唯一人类染色体的小鼠-人体细胞杂交系构建。含有人DNA插入片段的单个重组噬菌体通过其与总人DNA序列的杂交和其不与总小鼠DNA序列杂交来鉴定。然后从14个这样的重组噬菌体中的每一个亚克隆无重复的人DNA片段。使用独立的体细胞杂交将所有14个亚克隆片段分配到21号染色体。十三个片段已被区域映射使用体细胞杂交含有人类21易位染色体。两个探针定位在21q21.2易位断裂点的近端，11个探针定位在该断裂点的远端，将它们置于21q21.2- 21 q22区域。用于筛选限制性片段长度多态性的七种探针中的一种在与来自无关个体的基因组DNA杂交时识别多态性DNA片段。这14个独特的探针为研究人类21号染色体的结构和功能以及研究唐氏综合征的分子生物学提供了有用的工具。
To isolate DNA sequences unique to chromosome 21 we have used a recombinant-DNA library, constructed from a mouse-human somatic-cell hybrid line containing chromosome 21 as the only human chromosome. Individual recombinant phage containing human DNA inserts were identified by their hybridization to total human DNA sequences and by their failure to hybridize to total mouse DNA sequences. A repeat-free human DNA fragment was then subcloned from each of 14 such recombinant phage. An independent somatic-cell hybrid was used to assign all 14 subcloned fragments to chromosome 21. Thirteen of the fragments have been regionally mapped using a somatic-cell hybrid containing a human 21 translocation chromosome. Two probes map proximal to the 21q21.2 translocation breakpoint, and 11 probes map distal to this breakpoint, placing them in the region 21q21.2-21q22. One of seven probes used to screen for restriction-fragment-length polymorphisms recognized polymorphic DNA fragments when hybridized to genomic DNA from unrelated individuals. These 14 unique probes provide useful tools for studying the structure and function of human chromosome 21 as well as for investigating the molecular biology of Down syndrome.