CONSTRUCTION OF A PHYSICAL MAP FOR HUMAN CHROMOSOME 3

人类3号染色体由大约2亿个碱基对组成 关于DNA的。许多特定的重排都会影响这条染色体 肺癌、肾癌、白血病、颅面等疾病 和其他发育异常，使这条染色体富含 生物感兴趣的区域。许多工具可帮助您深入了解 对这条染色体的分析已经展开。这些措施包括： (1)广泛的体细胞杂交克隆面板，允许 准确、快速的分子探针区域定位；(2) 多个完整的Lambda和粘粒基因组文库 染色体及其区域；(3)分离 在来自这些文库的大约170个独特的序列探针中， 包括36个包含多个稀有宇宙的“星团宇宙” 限制性内切酶切割位点及其交叉杂交 对多个物种的DNA具有高频率，以及(4) 脉冲场电泳技术的改进 促进分析并将解析范围扩大到 大约1000万个碱基对。 考虑到这一点，我们建议：(1)隔离和区域分配 至少200个来自3号染色体的唯一序列探针。如果 随机分布的这将提供一个分子探针 每1000 kb的DNA。(2)绘制自然地图。 用这些探针和脉冲场电泳法进行染色体分析。 (3)鉴定常见的限制性片段长度多态 (RFLP)使用这些探头。遗传连锁分析将是 与其他调查人员合作完成。

Human chromosome 3 consists of approximately 200 million base pairs of DNA. Many specific rearrangements affect this chromosome in disorders such as lung and kidney cancer, leukemia, craniofacial and other developmental anomalies, making this chromosome a rich area of biologic interest. Many tools to begin an in depth analysis of this chromosome have been developed. These include: (1) an extensive somatic cell hybrid clone panel which permits the accurate and rapid regional mapping of molecular probes; (2) multiple lambda and cosmid genomic libraries of both the intact chromosome as well as regions thereof; (3) isolation of approximately 170 unique sequence probes from these libraries, including 36 "cluster cosmids" which contain multiple rare restriction endonuclease cleavage sites and which cross-hybridize with high frequency to the DNA of multiple species, and (4) improvements in pulsed-field electrophoretic technology which facilitate the analysis and expand the limits of resolution to approximately 10 million base pairs. With this in mind we propose: (1) To isolate and regionally assign a minimum of 200 unique sequence probes from chromosome 3. If randomly distributed this would provide a molecular probe for every 1000 Kb of DNA. (2) To construct a physical map of the chromosome using these probes and pulsed-field electrophoresis. (3) To identify common restriction fragment length polymorphisms (RFLP's) using these probes. The genetic linkage analysis will be done in collaboration with other investigators.