Systematic generation of sequence-tagged sites for physical mapping of human chromosomes: application to the mapping of human chromosome 7 using yeast artificial chromosomes.
Systematic generation of sequence-tagged sites for physical mapping of human chromosomes: application to the mapping of human chromosome 7 using yeast artificial chromosomes.
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系统生成用于人类染色体物理作图的序列标记位点：应用酵母人工染色体进行人类 7 号染色体作图。
DOI：
10.1016/0888-7543(91)90062-j
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发表时间：
1991
期刊：
影响因子：
4.4
通讯作者：
Olson,MV
中科院分区：
文献类型：
--
作者：
Green,ED;Mohr,RM;Idol,JR;Jones,M;Buckingham,JM;Deaven,LL;Moyzis,RK;Olson,MV
DNA 长程物理图谱开发的基础是重组克隆内标志物的检测和定位。序列标记位点 (STS) 是可以通过聚合酶链式反应 (PCR) 特异性检测的短 DNA 片段，可以用作此类标志。我们的兴趣是通过在酵母人工染色体 (YAC) 克隆中定位 STS 来构建整个人类染色体的物理图谱。在这里，我们描述了系统生成大量针对人类 7 号染色体的 STS 的通用策略。这些 STS 可以通过对 7 号染色体 DNA 的匿名片段进行测序后开发的 PCR 检测来检测，该匿名片段源自流式分选染色体或由人仓鼠杂交细胞系 DNA 制成的 lambda 克隆。我们的 STS 生成方法专为开发能够筛选大型 YAC 文库的 PCR 测定而定制。在这项研究中，我们报告了 100 个针对人类 7 号染色体的新 STS 的生成。
Basic to the development of long-range physical maps of DNA are the detection and localization of landmarks within recombinant clones. Sequence-tagged sites (STSs), which are short stretches of DNA that can be specifically detected by the polymerase chain reaction (PCR), can be used as such landmarks. Our interest is to construct physical maps of whole human chromosomes by localizing STSs within yeast artificial chromosome (YAC) clones. Here we deacribe a generalized strategy for the systematic generation of large numbers of STSs specific for human chromosome 7. These STSs can be detected by PCR assays developed following the sequencing of anonymous pieces of chromosome 7 DNA, which was derived from flow-sorted chromosomes or from lambda clones made from DNA of a human-hamster hybrid cell line. Our approach for STS generation is tailored for the development of PCR assays capable of screening a large YAC library. In this study, we report the generation of 100 new STSs specific to human chromosome 7.