Isolation and chromosomal localization of unique DNA sequences from a human genomic library.
Isolation and chromosomal localization of unique DNA sequences from a human genomic library.
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从人类基因组库中分离和染色体定位独特的 DNA 序列。
DOI：
10.1073/pnas.79.3.865
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发表时间：
1982
影响因子：
11.1
通讯作者：
Davidson,JN
中科院分区：
文献类型：
--
作者：
Kao,FT;Hartz,JA;Law,ML;Davidson,JN
筛选来自人类基因组文库的重组噬菌体，以鉴定仅具有独特序列的人类 DNA 插入片段。在大约 1% 的筛选噬菌体中发现了独特的人类插入片段。对一种重组噬菌体 P3-2 进行了详细研究。它包含 14.7 KB 的人类插入片段，具有四个内部 EcoRI 切割位点。构建了 EcoRI 和 BamHI 位点的限制性图谱。 32P 标记的 P3-2 探针与 EcoRI 消化的总人类 DNA 的 Southern 印迹杂交，在与 P3-2 中包含的人类插入片段相对应的位置产生了不同的条带。通过使用一系列含有独特人类染色体组合的人-中国仓鼠体细胞杂交体，通过印迹杂交和同线性分析，噬菌体P3-2中的人类DNA片段被分配到人类22号染色体。此外，噬菌体 P3-10 中的另一个人类 DNA 片段（11.4 KB）通过类似的程序被分配给人类 10 号染色体。通过这种方法，可以分离出更多独特的DNA序列，分配给特定的人类染色体，并用作人类基因组中基因作图和连锁、多态性和其他遗传学研究的遗传标记。
Recombinant bacteriophage lambda from a human genomic library were screened to indentify human DNA inserts having only unique sequences. Unique human inserts were found in about 1% of the phage screened. One recombinant phage, P3-2, was studied in detail. It contains a human insert of 14.7 kilobases with four internal EcoRI cleavage sites. A restriction map was constructed for EcoRI and BamHI sites. Hybridization of the 32P-labeled P3-2 probe to a Southern blot of EcoRI-digested total human DNA yielded distinct bands at positions corresponding to the human insert fragments contained in P3-2. By using a series of human-Chinese hamster somatic cell hybrids containing unique combinations of human chromosomes, the human DNA segment in phage P3-2 was assigned to human chromosome 22 by blot hybridization and synteny analysis. In addition, another human DNA segment, 11.4 kilobases, in phage P3-10 was assigned to human chromosome 10 by similar procedures. With this approach, more unique DNA sequences can be isolated, assigned to specific human chromosomes, and used as genetic markers for gene mapping and linkage, polymorphism, and other genetic studies in the human genome.