Kunkel L M, Tantravahi U, Eisenhard M, Latt S A
Nucleic Acids Res. 1982 Mar 11;10(5):1557-78. doi: 10.1093/nar/10.5.1557.
Fluorescence activated sorting of chromosomes from 49,XXXXY human lymphoblasts has been used to obtain DNA enriched for the human X. This DNA was cloned in lambda phage Charon 21A to obtain a library of approximately 60,000 pfu. Phage inserts free of human highly repeated DNA sequences are localized to different regions of the human X by two independent hybridization analyses. The first utilized comparative hybridization to rodent-human hybrid cell DNA samples containing all or known portions of the human X, while the second was based on hybridization dosage to DNA samples from human cell lines differing in the number of X chromosomes or X chromosome segments. Of five unique sequence inserts tested, three were X chromosome specific and were localized to regions Xpter leads to Xcen, Xql leads to Xq22 and Xq24 leads to Xqter, respectively. The library presented here represents a highly enriched source of human X chromosome-specific DNA sequences.
利用荧光激活分选技术从49,XXXXY人类淋巴母细胞中分离染色体，以获取富含人类X染色体的DNA。该DNA被克隆到λ噬菌体Charon 21A中，构建了一个约60,000个噬菌斑形成单位（pfu）的文库。通过两种独立的杂交分析，将不含人类高度重复DNA序列的噬菌体插入片段定位到人类X染色体的不同区域。第一种方法是与含有全部或已知部分人类X染色体的啮齿动物-人类杂交细胞DNA样本进行比较杂交，第二种方法是基于对来自X染色体数量或X染色体片段数量不同的人类细胞系DNA样本的杂交剂量分析。在测试的五个独特序列插入片段中，三个是X染色体特异性的，分别定位于Xpter至Xcen、Xq1至Xq22和Xq24至Xqter区域。本文提供的文库代表了人类X染色体特异性DNA序列的高度富集来源。
