Kao F T, Hartz J A, Law M L, Davidson J N
Proc Natl Acad Sci U S A. 1982 Feb;79(3):865-9. doi: 10.1073/pnas.79.3.865.
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.
对来自人类基因组文库的重组噬菌体λ进行筛选，以鉴定仅具有独特序列的人类DNA插入片段。在大约1%被筛选的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。它包含一个14.7千碱基的人类插入片段，有四个内部EcoRI切割位点。构建了EcoRI和BamHI位点的限制性图谱。用32P标记的P3 - 2探针与经EcoRI消化的人类总DNA的Southern印迹杂交，在与P3 - 2中所含人类插入片段相对应的位置产生了明显的条带。通过使用一系列含有独特人类染色体组合的人 - 中国仓鼠体细胞杂种，通过印迹杂交和同线分析将噬菌体P3 - 2中的人类DNA片段定位到人类22号染色体上。此外，通过类似的方法将噬菌体P3 - 10中的另一个11.4千碱基的人类DNA片段定位到人类10号染色体上。通过这种方法，可以分离出更多独特的DNA序列，将其定位到特定的人类染色体上，并用作人类基因组中基因图谱绘制、连锁分析、多态性及其他遗传研究的遗传标记。
