MOLECULAR ANALYSIS OF HUMAN CHROMOSOME 21 AND 3

人类3号染色体的特异性重排与 恶性肿瘤如小细胞肺癌，肾细胞癌， 白血病和几种发育障碍 人类21号染色体是 与唐氏综合症、白血病和阿尔茨海默病有关。 这两 染色体含有大约2亿和5千万个碱基对， DNA，因此总共可能含有4000个 8,000个基因 总共只有不到100个基因被发现， 这意味着许多有价值的生物信息不容易获得。 为了增加我们对染色体组织的了解， 分离生物学上重要的基因，这项建议的具体目标 构建酵母人工染色体（YAC）文库， 3号和21号染色体的组成图。 成分图将 提供基因丰富区域的信息，并将允许克隆 以及其他不寻常的染色体区域。 YAC图书馆将 用放置在脉冲场上的独特序列进行筛选 图，以及来自各种细胞系和组织的cDNA。 这将导致 在i）YAC克隆的快速区域定位，ii）鉴定 用于脉冲场连接的另外的独特序列，iii）产生 用于寻找染色体易位断裂点的其它探针， 和iv）鉴定表达的序列，包括在所述表达序列中的一些序列， 易位断点附近。 长期目标包括：i） 受AML-M2中所见的8; 21断裂点影响的基因的表征， 以及其他人，二）发展一系列的大型 插入克隆以跨越大的染色体区域，iii）产生 用于转基因小鼠研究的良好表征的YAC。

Specific rearrangements of human chromosome 3 are associated with malignancies such as small cell lung cancer, renal cell carcinoma, leukemia, and several developmental disorders. Human chromosome 21 is associated with Down Syndrome, leukemia and Alzheimer's Disease. These two chromosome contain approximately 200 million and 50 million base pairs of DNA, respectively, and therefore together potentially contain between 4,000 and 8,000 genes. Fewer than 100 genes in total have been identified, meaning that much valuable biological information is not easily accessible. In order to increase our understanding of chromosomal organization and to isolate biologically important genes, the specific aims of this proposal are to construct yeast artificial chromosome (YAC) libraries and compositional maps for chromosomes 3 and 21. The compositional maps will provide information on gene rich regions and will allow the cloning of these and other unusual chromosomal regions. The YAC libraries will be screened with unique sequences that have been placed on the pulsed field maps, and with cDNAs from various cell lines and tissues. This will result in i) rapid regional localization of YAC clones, ii) identification of additional unique sequences for pulsed field linkage, iii) generation of additional probes for the search for chromosomal translocation breakpoints, and iv) identification of expressed sequences, including some in the vicinity of translocation breakpoints. Long term objectives include: i) characterization of genes affected by the 8; 21 breakpoint seen in AML-M2, as well as those from others, ii) the development of a series of large insert clones to span large chromosomal regions, iii) the generation of well characterized YACs useful for transgenic mouse studies.