GENOMICS RESOURCES AND INFRASTRUCTURE FOR THE ZEBRAFISH

斑马鱼最近成为研究早期脊椎动物个体发育的重要模型，目前通过诱变筛选鉴定的发育突变体数量超过2000种。负责这些突变表型的几个基因可能具有人类对应物，其可能引起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，都必须对这些基因进行分子“克隆”。虽然到目前为止，已经克隆了一些这样的斑马鱼基因，但用于启动定位克隆项目和染色体步移的基因组学基础设施还没有很好地建立，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此类项目的负担感到气馁。此外，由于这一承诺所需的时间和财政资源是相当大的，生物学家是进一步远离解决关键和有意义的问题密切相关的发育生物学的更深入的理解。正如trans-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，以促进突变基因的识别是该社区的核心任务。因此，这项建议的首要目标是产生和维持大插入基因组学资源，这些资源是斑马鱼社区容易获得的，以便更迅速地克隆发育相关基因。这将通过从斑马鱼近交系构建改进的BAC和PAC文库来实现，产生这些文库以及现有的大插入文库的池，向社区分发基因组学试剂（沿着适当的用户友好的协议），并作为基因组资源、信息和专业知识的集中储存库。与这些资源有关的数据将存储在ZFIN中，并将开发适当的输入脚本。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，这样产生的文库将成为未来大规模斑马鱼基因组物理作图和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.