INTEGRATED ZEBRAFISH GENOMIC RESOURCES

描述：（申请人的摘要）斑马鱼Danio rerio是一种 用于发育突变分析重要脊椎动物模型系统 流程.斑马鱼的有效利用需要各种 尚未充分开发的遗传和基因组资源，包括 序列标签（EST），高密度遗传和物理，以及强大的近交系 绘制菌株图谱。我们提出了一个整合基因组资源的方法 开发，包括EST测序，将用于开发 序列标记位点（STS），这反过来将用于识别单个 近交系之间的核苷酸多态性（SNP），并绘制在 辐射杂种（RH）和减数分裂作图板。 (1)表达序列标签（EST）的产生，从不同阶段的 斑马鱼发育。我们将生成5'和3'序列， 来自50，000个不同簇中的每一个的代表性cDNA， 寡核苷酸杂交指纹分析278，000独立 斑马鱼cDNA克隆。 (2)STS开发和辐射混合映射。我们将创造10000个 来自3' EST读数的STS标记和辐射上的基因型5，000个STS 用于构建标记密集物理图的混合面板，包括具有 SNP允许整合物理和遗传图谱。 (3)单核苷酸多态性标记开发和减数分裂作图。 我们将重新测序10,000个基于EST的STS，这些STS来自用于 减数分裂作图以鉴定单核苷酸多态性。我们将类型 减数分裂作图板上限制酶位点的1800个SNP， 标记密集遗传图谱的生成。 用EST鉴定多达50，000个人类基因的斑马鱼同源物 测序将有助于这些基因的功能分析， 在人类身上。此外，10，000条斑马鱼EST的遗传和物理作图 将为斑马鱼突变提供候选基因， 启动定位克隆项目最后，地图比较 斑马鱼和人类基因组的位置。这将使斑马鱼 突变来揭示人类基因的作用， 人类疾病的基础。

DESCRIPTION: (Applicant's abstract) The zebrafish Danio rerio is an important vertebrate model system for mutational analysis of developmental processes. Efficient utilization of the zebrafish requires a variety of genetic and genomic resources not yet fully developed, including expressed sequence tags (ESTs), high density genetic and physical, and robust inbred mapping strains. We propose an integrated approach to genomic resource development, including EST sequencing, that will be used to develop sequence tagged sites (STSs), that will in turn be used to identify single nucleotide polymorphisms (SNPs) between inbred strains, and mapped on radiation hybrid (RH) and meiotic mapping panels. (1) Generation of Expressed Sequence Tags (ESTs) from various stages of zebrafish development. We will generate 5' and 3' sequences from representative cDNAs from each of 50,000 different clusters identified by oligonucleotide hybridization fingerprint analysis of 278,000 independent zebrafish cDNA clones. (2) STS development and Radiation Hybrid mapping. We will generate 10,000 STS markers from 3' EST reads and genotype 5,000 STSs on a Radiation Hybrid panel to construct a marker dense physical map, including STSs with SNPs to allow integration of physical and genetic maps. (3) Single nucleotide polymorphism marker development and Meiotic mapping. We will re-sequence 10,000 EST-based STS from inbred strains used in meiotic mapping to identify Single Nucleotide Polymorphisms. We will type 1800 SNPs at restriction enzyme sites on meiotic mapping panels for generation of a marker dense genetic map. Identification of up to 50,000 zebrafish homologues of human genes by EST sequencing will facilitate functional analyzes of these genes not possible in humans. Moreover, genetic and physical mapping of 10,000 zebrafish EST will provide candidate genes for zebrafish mutations and a dense framework for initiating positional cloning projects. Finally, comparison of map positions of zebrafish and human genomes. This will allow zebrafish mutations to reveal roles for human genes and increase our understanding of the basis of human disease.