HIGH RESOLUTION PHYSICAL MAPS OF 4P AND 5Q

这些研究的长期目标是利用来自高密度脂蛋白的信息 人体选定区域的分辨率、远程物理地图 基因组以了解遗传的单基因和 并帮助理解染色体的功能组织 人类基因组。该提案的第一部分涉及测试和 实施两种新方法以促进远程物理 映射。第一种技术涉及插入逆转录病毒载体。 具有极其罕见的(10碱基和12碱基)限制性内切酶 进入人类染色体的识别位点，然后将用于开发 一步完成5-10Mb的物理地图。第二项新技术是 在以E.Coli为基础的克隆系统中进行，该系统允许克隆非常 细菌宿主中的大(&gt；150 kb)外源DNA片段。这些新的 与现有重组DNA和体细胞遗传学相结合的方法 这些技术将被用来开发高分辨率、远程物理 基因组的两个特定区域的图谱。第一个区域是5q21-5q23， 家族性腺瘤性息肉病基因周围区域 冒号(APC)。该基因突变导致结肠癌的易感性 癌症和疾病是作为常染色体显性遗传的 接近100%的穿透力。要映射的另一个区域，它也将 作为检测大片段大肠杆菌克隆载体的系统， 是4号染色体短臂的远端部分。这个区域 包含几个具有医疗价值的地区，包括亨廷顿 4P或Wolf-Hirschhorn的致病基因和临界区 综合症，以及其他几个有趣的特征，如一个大的 多个VNTR序列；至少一个重组热点；以及多个 汉能薄膜发电群岛。除了回答有关这些问题的具体问题外 人类基因组的两个区域，有待开发和使用的方法 这个项目应该有助于绘制基因组的其他部分。

The long term goal of these studies is to use information derived from high resolution, long-range physical maps of selected regions of the human genome to understand the molecular basis of inherited single gene and chromosomal disorders and to help understand the functional organization of the human genome. The first part of this proposal involves the testing and implementation of two new approaches to facilitate long range physical mapping. The first technique involves the insertion of retroviral vectors with extremely rare (10 base and 12 base) restriction endonuclease recognition sites into human chromosomes which will then be used to develop physical maps of 5-10Mb in one step. The second new technique to be pursued in an E. coli-based cloning system which allows the cloning of very large (>150 Kb) exogenous DNA fragments in the bacterial host. These new approaches together with existing recombinant DNA and somatic cell genetic techniques will be used to develop high resolution, long range physical maps of two specific regions of the genome. The first area is 5q21-5q23, the region surrounding the gene for familial adenomatous polyposis of the colon (APC). Mutations in this locus result in predisposition to colon cancer and the disease is inherited as an autosomal dominant trait with close to 100% penetrance. The other region to be mapped, which will also serve as the system for testing the large fragment E. coli cloning vector, is the distal portion of the short arm of chromosome 4. This region contains several regions of medical interest, including the Huntingtons disease gene and the critical region for the 4p- or Wolf-Hirschhorn syndrome, as well as several other interesting features such as a large number of VNTR sequences; at least one recombination hot spot; and a number of HTF islands. In addition to answering specific questions about these two regions of the human genome, the approaches to be developed and used in this project should aid in mapping other portions of the genome.