DETAILED MAPPING AND RECOMBINATION OF CHROMOSOME FOUR

本研究的主要目的是分离、研究和定位dna。 人类4号染色体的片段。提出的实验将 分为两个领域，将同时进行调查。 首先，使用酵母人工染色体(YAC)的方法 克隆人类DNA的大片段将作为 4号染色体的分离.人类序列的表示 在克隆的YAC池中，特别注意重复 序列(VNR、LINE、SINE和ALPHOID重复)将是 检查过了。此外，长期的稳定性和对 野生型酵母和突变体中的重排 重组和DNA修复基因的改变将是 下定决心。据报道人类片段可刺激(“热点”)或 抑制“冷点”)的重组也将被研究 对酵母重组的影响。第二，对小说的研究 一种称为连锁缺失的克隆DNA序列排序方法 均衡图谱将得到扩展。在建立1%的 4号染色体的遗传连锁图谱(通过传统方法)， 不平衡映射将应用于定位的有序片段 相隔不到1厘米。这项技术将成为一种强大的 物理测绘程序的替代(例如，脉冲场凝胶 电泳法(PFGE))正是在这些区域中 难以物理解决--1%重组的冷点 代表107个或更多BP。不平衡映射取决于 重组距离，并在1%的地区同样有效 重组等于107个碱基对，因为在1%等于105个碱基的区域。 然后，该项目将生成几种形式的完整地图 四号染色体和酵母中四号染色体的完整文库。 直接利用染色体图谱进行重组的研究 四是通过研究特定的行为来间接地 酵母重组系统中的四号染色体序列将是 制造。该项目将在以下方面提供重要步骤 建立、使用不平衡作为工具来确认或 扩展物理地图。最后，它将提供详细的见解 探讨酵母菌和人类的重组机制-- 产生种群多样性的基本机制。

The major aim of this research is to isolate, study and map DNA fragments from human chromosome 4. The experiments proposed will fall into two areas which will be investigated simultaneously. First, the method of using yeast artificial chromosomes (YACs) to clone large segments of human DNA will be studied as part of the isolation of chromosome 4. The representation of human sequences in the cloned YAC pool, paying special attention to repeated sequences (VNTRs, LINES, SINES and alphoid repeats) will be examined. In addition, the long term stability and resistance to rearrangement in both wild type yeast and in mutants with alterations in recombination and DNA repair genes will be determined. Human fragments reported to stimulate ("hotspots") or depress "coldspots") recombination will also be studied for their effects on recombination in yeast. Second, studies of a novel method for ordering cloned DNA sequences called linkage dis- equilibrium mapping will be expanded. After establishing a 1% genetic linkage map (by traditional methods) for chromosome 4, disequilibrium mapping will be applied to order fragments located less than lcM apart. This technique will serve as a powerful alternative to physical mapping procedures (e.g., pulse field gel electrophoresis (PFGE)) in exactly those regions that are most difficult to physically resolve--coldspots where 1% recombination represents 107 or more bp. Disequilibrium mapping depends upon recombination distances and is as effective in a region where 1% recombination equals 107 bp as in a region where 1% equals 105 bp. The project will then generate several forms of complete maps of chromosome four and a complete library of chromosome four in yeast. Studies of recombination both directly using the maps of chromosome four and indirectly by studying the behavior of particular chromosome four sequences in a yeast recombination system will be made. This project will provide an essential step in establishing,the use of disequilibrium as a tool to confirm or extend physical maps. Finally, it will provide detailed insights into the mechanisms of recombination in yeast and man - one of the basic mechanisms of generating population diversity.