CHROMOSOME 5 SPECIFIC CDNA'S

在这个项目中，我们将产生和评估三个染色体5- 通过直接选择特异性cDNA文库： 能够分离甚至低丰度的cDNA。 选择的cDNA将 用于筛选一组排列的5号染色体特异性 基因组克隆，从而鉴定至少一些转录的 基因组区域。 cDNA也将被构建成一个排列整齐的高- 将用基因组YAC筛选的50，000个克隆的密度集， 粘粒重叠群，因为它们被开发，从而增加表达数据到 扩展物理图谱，直接鉴定表达的cDNA， 通过基因组重叠群。 500个5号染色体特异性选择的cDNA将被克隆。 转换为表达序列标签（EST），放置在物理图谱上 使用辐射杂交作图，并用于确定其同源 基因组序列 使用这些方法，我们将能够注释 粘粒、YAC和辐射混合物理图， 基因，并能够将各种地图锚在一起。 的应用 多重选择和cDNA标准化策略也应该证明 可用于组织特异性转录图谱的快速推导 在人类基因组的大部分区域， 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.