GENOMICS RESOURCES AND INFRASTRUCTURE FOR THE ZEBRAFISH

斑马鱼（Danio ior）最近成为研究早期脊椎动物个体发育的重要模型，通过诱变筛选发现的发育突变体数量目前超过2000个。导致这些突变表型的几个基因可能在人类中有对应的基因，这可能会导致相应的异常状况和出生缺陷。为了研究这些发育过程的潜在生物学，无论是单个的还是全部的，这些基因被分子“克隆”是一个必要的条件。虽然迄今为止已经克隆了一些这样的斑马鱼基因，但启动位置克隆项目和染色体行走的基因组学基础设施还没有很好地建立起来，特别是关于大插入克隆试剂。许多发育生物学家仍然对重新启动这类项目的负担感到畏惧。此外，由于这一承诺所需的时间和财政资源是相当可观的，生物学家离解决与更深入地理解发育生物学相关的关键和有意义的问题越来越远。正如跨nih斑马鱼协调委员会所建议的那样，更好的基因组学基础设施促进突变基因的鉴定是该社区的当务之急。因此，本提案的首要目标是生成和维护大插入基因组资源，以便更快速地克隆与发育相关的基因，以便于斑马鱼群落容易获得。这将通过从近交系的斑马鱼中构建改进的BAC和PAC文库，生成这些文库以及现有的大插入文库的库，向社区分发基因组学试剂（以及适当的用户友好协议），并作为基因组资源，信息和专业知识的集中存储库来实现。关于这些资源所产生的数据将存储在ZFIN中，并将为它们的输入编写适当的脚本。传播实物和虚拟资源将鼓励社区内多层次的方案间合作。此外，由此产生的文库将成为未来大规模物理制图和斑马鱼基因组测序以及功能基因组学实验的组成部分。

The zebrafish, Danio rerior, has recently emerged as an important model for studying early vertebrate ontogeny, the number of developmental mutants identified via mutagenesis screens currently exceeding 2,000. Several of the genes responsible for these mutant phenotypes will likely have human counterparts which may give rise to corresponding aberrant conditions and birth defects. In order to study the underlying biology o these developmental processes either singly or in tot or in toto, it is a conditio sine qua non that these genes be molecularly "cloned". While to date a few such zebrafish genes have been cloned, the genomics infrastructure for initiating positional cloning projects and chromosomal walks is not well established, particularly with regard to large-insert cloning reagents. Many developmental biologists remain daunted by the burden of initiating such projects afresh. Additionally, since the time and financial resources required for this commitment are considerable, the biologist is distanced further from addressing critical and meaningful questions germane to a more in-depth understanding of the developmental biology. As recommended by the trans-NIH zebrafish coordinating committee, better genomics infrastructure for facilitating the identification of mutant genes is a central imperative for the community. Thus, the overriding objective of this proposal is to generate and maintain large-insert genomics resources that are readily accessible to the zebrafish community in order to more expeditiously clone developmentally relevant genes. This will be accomplished by constructed improved BAC and PAC libraries from inbred strains of zebrafish, generating pools of these libraries as well as from an existing large-insert library, distributing genomics reagents (along with appropriate user-friendly protocols) to the community, and serving as a centralized repository for genome resources, information and expertise. Data generated with respect to these resources will be stored in ZFIN, and proper scripts for their entry will be developed. Dissemination of resources, both physical and virtual, will encourage, multi-tiered, inter-programmatic cooperation within the community. In addition, the libraries so generated will be an integral component for future large-scale physical mapping and sequencing of the zebrafish genome, as well as functional genomics experiments.