HUMAN DISEASE ISOLATION USING GENETIC LINKAGE AND MOLECULAR CLONING TECHNIQUES

这项研究的长期目标是开发一种通用的 用于分离对应于遗传修饰的任何人类基因的方法 病源 最终，这些方法应该允许 分离出大量的基因， 纤维化，躁狂抑郁症，肝豆状核变性，多囊肾 疾病和其他类似的疾病。 一旦基因 已确定的确切遗传和生化性质的 疾病可以确定。 从这一理解， 可以进行有针对性的调查，以便及早发现和及早治疗。 治疗 一般的方法是使用最近开发的 限制性片段长度多态性（RFLP）图谱 首先定位一个致病位点，然后我们将开发一种新的酵母菌 人工染色体载体，用于克隆两个 侧翼RFLP。 然后，疾病位点将被包含在 克隆DNA DNA的数量大约在100万到500万之间 碱基对（Mb），并可能包含一些正常的 正常基因和缺陷基因 以限缩检索 缺陷基因的高分辨率RFLP图谱将由 一种新的物理作图方法和精细结构重组 并通过调查绘制连锁不平衡图 家庭传播疾病。 从这张高分辨率地图上 基因应该被精确定位 信使RNA，s将是 在这个地区进行调查，确切的基因将由 直接DNA测序。

The long term objective of this research is to develop a general approach for isolating any human gene corresponding to a genetic disease locus. Ultimately these approaches should allow the isolation of large numbers of genes including those for cystic fibrosis, manic depression, Wilson's disease, Polysistic Kidney disease and a host of other similar diseases. Once the genes have been identified the exact genetic and biochemical nature of the disease can be determined. From this understanding a much more directed investigation can be made for early detection and early treatment. The general approach is to use the recently develop restriction fragment length polymorphism (RFLP) map of the human to first locate a disease locus, we then will develop a new yeast artificial chromosome vector for cloning the DNA between two flanking RFLP. The disease locus will then be contained within the cloned DNA. The amount of DNA will approximately 1 to 5 million basepairs (Mb) and will probably contain a number of normal of normal genes with the defective gene. To narrow the search defective gene a high resolution RFLP map will be generated by a new physical mapping procedure an a fine structure recombination and linkage disequilibrium map will be prepared by surveying families transmitting the disease. From this high resolution map the gene should be closely pinpointed. Messenger RNA,s will be surveyed in this area and the exact gene will be determined by direct DNA sequencing.