CONSTRUCTION OF A PHYSICAL MAP FOR HUMAN CHROMOSOME 3

人类 3 号染色体物理图谱的构建

• 批准号: 3298780
• 负责人: HARRY A. DRABKIN
• 金额: $ 16.53万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-09-29 至 1991-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3298780
• 关键词: DNA; gel electrophoresis; genetic library; genetic mapping; genetic markers; genome; human tissue; hybrid cells; linkage mapping; molecular cloning; nucleic acid probes; plasmids

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.

人类3号染色体由大约2亿个碱基对组成 的DNA。 许多特定的重排影响这条染色体， 例如肺癌和肾癌、白血病、颅面神经疾病 和其他发育异常，使这条染色体成为一个丰富的 生物感兴趣的领域。 许多工具可以开始深入研究 对这种染色体的分析已经被开发出来。 其中包括： (1)一个广泛的体细胞杂交克隆面板，允许 分子探针的准确和快速区域定位;（2） 多个λ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个独特序列探针中， 包括36个“集群宇宙”，其中包含多种罕见的 限制性内切酶切割位点，并且其交叉杂交 与多个物种的DNA的高频率，和（4） 脉冲场电泳技术的改进 促进分析并扩大分辨率的范围， 大约一千万个碱基对 为此，我们建议：（1）分离和区域分配 最少200个来自3号染色体的独特序列探针。 如果 随机分布，这将提供一种分子探针， 每1000 Kb的DNA。 (2)为了构建一个物理地图， 染色体使用这些探针和脉冲场电泳。 (3)鉴定常见的限制性片段长度多态性 (RFLP的）使用这些探针。 遗传连锁分析将是 与其他研究人员合作。