Approaches to physical mapping of the human genome.
Approaches to physical mapping of the human genome.
复制标题
人类基因组物理绘图的方法。
DOI：
10.1101/sqb.1986.051.01.014
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发表时间：
1986
期刊：
影响因子：
--
通讯作者：
Cantor,CR
中科院分区：
文献类型：
--
作者：
Smith,CL;Cantor,CR
研究人类基因组图谱的宏观方法包括细胞遗传学、体细胞遗传学和连锁分析。这些方法中的每一种都是强大的，并且都是互补的。然而，在实践中，当结构数据需要比10，000 kb更精细的分辨率时，每一个都变得越来越乏味。在不久的将来，这些方法中的任何一种似乎都不可能扩展到以1000 kb的分辨率提供人类基因组的常规分析。检查人类基因组图谱的分子方法包括克隆、限制性酶切图谱和DNA片段测序。实际上，用限制性酶切图谱分析10- 50 kb的区域几乎是微不足道的。通过染色体步移技术，可以将这种图谱扩展到几百种酶。然而，随着区域大小的增加以及DNA片段被高度重复的序列所支配，这些变得相当乏味。虽然1000 kb的步移是可行的，但除非特定的DNA区域令人感兴趣，否则很难以极大的热情来完成这样的任务。为了连接分子和宏观技术，需要的是一种构建具有100至1000 kb分辨率的物理图谱的方法。在这篇文章中，我们描述了四种技术，一起，允许这样一个粗糙的限制性图谱的建设。每一种技术都是在简单的生物体如酵母或细菌上开发或测试的，但现在每一种技术都被证明是可行的，可用于对人类样本的可比研究。我们还概述了一种策略，似乎是一个相当有效的方式来应用这些技术来构建每个人类染色体的完整的物理图谱。最后，我们简要讨论了这些地图的效用。
Macroscopic methods of examining the map of the human genome include cytogenetics, somatic cell genetics, and linkage analysis. Each of these methods is powerful, and all are quite complementary. In practice, however, each becomes progressively more tedious when structural data are required at finer resolution than 10,000 kb. It seems unlikely that any of these methods will be extended, in the near future, to provide routine analysis of the human genome at 1000-kb resolution.Molecular methods of examining the map of the human genome involve cloning, restriction mapping, and sequencing DNA fragments. In practice the analysis of a 10-to 50-kb region by restriction mapping is nearly trivial. The extension of such maps to several hundred kilobases is possible by chromosome-walking techniques. However, these become quite tedious as the size of the region increases and as segments of DNA become dominated by highly repeated sequences. Although walks of 1000 kb are practical, it would be hard to approach such a task with great enthusiasm unless the particular DNA region were of compelling interest. To bridge the molecular and macroscopic techniques, what is needed is a way to construct a physical map with 100-to 1000-kb resolution. In this article, we describe four techniques that, together, allow the construction of such a coarse restriction map. Each technique was developed or tested on simple organisms like yeast or bacteria, but each has now been shown to be feasible for comparable studies on human samples. We also outline a strategy that appears to be quite an efficient way to apply these techniques to construct complete physical maps of each human chromosome. Finally, we briefly discuss the utility of such maps.