CHROMOSOME 5 SPECIFIC CDNA'S

在这个项目中，我们将生成并评估三条5号染色体- 直接选择特异的cDNA库：一种 能够分离出即使是低丰度的cDNA。选定的cDNA将 用于筛选一组特定于5号染色体的阵列 基因组克隆，从而识别至少部分转录的 基因组区域。这些DNA也将被构建成一个排列的高音- 50,000个克隆的密度集，将通过基因组YAC和 粘粒重叠群，从而将表达式数据添加到 扩展物理图谱并直接识别所表达的cDNA 通过基因组重叠群。500个特定于5号染色体的cDNA将被 转换为表达序列标签(EST)，放置在物理地图上 利用辐射杂交作图，并用于鉴定它们的同源 基因组序列。使用这些方法，我们将能够注释 COSMID、YAC和辐射混合物理贴图具有大量 基因，并能够将各种图谱锚定在一起。的应用 多重选择和cdna标准化策略也应该证明 在快速获得组织特异性转录图谱中有用 在人类基因组的大范围区域和在评估 编码序列在整个5号染色体上的分布。

In this project, we shall generate and evaluate three chromosome 5- specific cDNA libraries by direct selection: a technique that is capable of isolating even low abundance cDNAs. The selected cDNAs will be used en masse to screen an arrayed set of chromosome 5-specific genomic clones, thus identifying at least some of the transcribed genomic regions. The cDNAs will also be built into an arrayed high- density set of 50,000 clones that will be screened with genomic YAC and cosmid contigs as they are developed, thus adding expression data to the expanding physical map and directly identifying cDNAs that are expressed by genomic contigs. 500 chromosome 5-specific selected cDNAs will be converted to Expressed Sequence Tags (ESTs), placed on the physical map using radiation hybrid mapping, and used to identify their homologous genomic sequences. Using these methods, we will be able to annotate the cosmid, YAC, and radiation hybrid physical maps with a large number of genes and be able to anchor the various maps together. Applications of multiplex selection and cDNA normalization strategies should also prove useful in the rapid derivation of tissue specific transcription maps across large regions of the human genome and in assessing the distribution of coding sequences throughout chromosome 5.