International Zebrafish Mutagenic Protein Trap

“国际斑马鱼蛋白质捕集联盟” 摘要/概要 斑马鱼（Danio rerio）是研究核心蛋白质的杰出的非哺乳类脊椎动物系统。 脊椎动物生物学和行为以及人类疾病的建模。收集分子 表征的突变体系已经成为线虫、蝇和小鼠领域的有力工具。基因组 方法已经有力地表征了核基因组和转录组。然而，基因组 在动态背景下和体内对蛋白质组的全部复杂性的评估仍然是未知的。的 斑马鱼提供了第一个以整个脊椎动物为模板的蛋白质组综合分析 基因组 我们表明，诱变蛋白陷阱基因破坏转座子（GBT）是有效的和可逆的损失- 用于探索传统领域的功能工具，如发育生物学（包括器官发生） 而且还开辟了新的选择，包括行为的遗传学，如尼古丁反应的生物学。 一组Cre重组酶可调控的突变斑马鱼等位基因提供了探索关键基因的机会。 在组织、器官和细胞水平上解决脊椎动物生物学的问题。该应用程序在 对“PAR-08-139用研究工具和技术加强斑马鱼研究（R 01）”的回应， 在开发这些可回复突变等位基因的大规模收集方面取得了完整的进展， 通过协调一致的国际努力，我们将在项目结束时提供以下内容： I. 1000个新的诱变蛋白陷阱GBT斑马鱼品系与标签表达的基本描述 将建立模式，并通过可搜索的在线数据库实时提供 这些线路通过马约诊所斑马鱼分发立即可用 核心设施。 二.我们将对这些品系进行分子分析，并对斑马鱼基因组进行后续注释 项目，以确定被困蛋白质在这个集合。我们将继续利用我们成功的下一个 产生序列技术，然后进行连锁分析，以分离和映射GBT插入在 斑马鱼基因组所有序列信息最初将通过zfishbook.org发布，然后 整合在ZFIN、Fishmap和其他斑马鱼基因组数据库中。 三.我们将定期与斑马鱼国际资源中心（ZIRC）同步此集合， 长期存档并分发给斑马鱼社区。我们将使用最近建立的2D 条形码库存系统与最先进的精子冷冻保存方法，以促进定期 向ZIRC运送新线路。 这个建议是为了完成这个分子特征的收集，在硅片目录和分销网络 斑马鱼这种杰出的非哺乳类脊椎动物的可逆突变。这些显性标记 突变株系也代表了用于大规模行为和其他表型评估的理想底物， 基因组作为更广泛的倡议的一部分，使用功能注释脊椎动物基因组， 斑马鱼

"International Zebrafish Protein Trap Consortium" Abstract/Summary The zebrafish (Danio rerio) is the preeminent non-mammalian vertebrate system for the study of core vertebrate biology and behavior and for the modeling of human disease. Collections of molecularly characterized mutant lines have been powerful enabling tools for the nematode, fly and mouse fields. Genomic approaches have robustly characterized the nuclear genome and transcriptome. However, the genomic assessment of the full complexity of the proteome in a dynamic context and in vivo is still largely unknown. The zebrafish offers the first comprehensive analysis of the proteome using as template an entire vertebrate genome. We show that mutagenic protein trap gene-breaking transposons (GBTs) are effective and revertible loss- of-function tools for exploring traditional areas such as the biology of development (including organogenesis) but also open up new options including the genetics of behavior such as the biology of the nicotine response. A panel of Cre recombinase regulatable mutant zebrafish alleles offers the opportunity to explore critical questions in vertebrate biology at the tissue, organ and cellular level of resolution. This application is in response to "PAR-08-139 Enhancing Zebrafish Research with Research Tools and Techniques (R01)" to complete progress on the development of a large-scale collection of these revertible mutant alleles for the zebrafish community through a coordinated international effort. We will deliver the following at project's end: I. 1000 new mutagenic protein trap GBT zebrafish lines with basic description of the tagged expression patterns will be established and made available in real-time through the searchable online database zfishbook.org. These lines are immediately available through distribution by the Mayo Clinic Zebrafish Core Facility. II. We will conduct molecular analyses of these lines and subsequent annotation on the zebrafish genome project to identify the trapped proteins in this collection. We will continue to use our successful next generation sequence techniques followed by linkage analysis to isolate and map GBT insertions on the zebrafish genome. All sequence information will be initially distributed through zfishbook.org and then integrated in ZFIN, Fishmap and other zebrafish genomic databases. III. We will regularly synchronize this collection with the Zebrafish International Resource Center (ZIRC) for long-term archival and distribution to the zebrafish community. We will use a recently established 2D barcoding inventory system with a state-of-the-art sperm cryopreservation approach to facilitate regular shipping of new lines to ZIRC. This proposal is to complete this molecularly characterized collection, in silico catalog and distribution network of revertible mutations for the preeminent non-mammalian vertebrate, the zebrafish. These dominantly marked mutants lines also represent an ideal substrate for large-scale behavioral and other phenotypic assessment of the genome as a part of the broader initiative to functionally annotate the vertebrate genome using the zebrafish.