CHROMOSOME 5 SPECIFIC CDNA'S

5 号染色体特异性 CDNA

• 批准号: 3735832
• 负责人: Michael Lovett
• 金额: --
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3735832
• 关键词: artificial chromosomes; chromosomes; complementary DNA; embryo /fetus cell /tissue; gene expression; genetic library; genetic mapping; human genetic material tag; human tissue; in situ hybridization; nucleic acid hybridization; nucleic acid sequence; polymerase chain reaction; radiotracer; sequence tagged sites; southern blotting

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.

在这个项目中，我们将生成和评估三个5-染色体 通过直接选择的特定cDNA库：一种技术 能够隔离低丰度cDNA。 选定的cDNA将会 被大批用于筛选一组染色体5特异性的染色体 基因组克隆，因此至少识别了一些转录 基因组区域。 cDNA也将内置在阵列的高 - 将使用基因组YAC筛选的50,000个克隆的密度集和 宇宙重叠群的开发，从而将表达数据添加到 扩展物理图并直接识别表达的cDNA 通过基因组重叠群。 500染色体5特异性选定的cDNA将是 转换为放置在物理图上的表达序列标签（ESTS） 使用辐射混合映射，并用于识别其同源 基因组序列。 使用这些方法，我们将能够注释 宇宙，YAC和辐射杂交物理图和大量 基因并能够将各种地图固定在一起。 申请 多重选择和cDNA归一化策略也应证明 在组织特异性转录图的快速推导中有用 在人类基因组的大区域以及评估 整个染色体5的编码序列的分布。