GENOMICS RESOURCES AND INFRASTRUCTURE FOR THE ZEBRAFISH

斑马鱼，Danio rerior，最近成为研究早期脊椎动物个体发育的重要模型，通过突变筛选确定的发育突变数量目前超过2000。导致这些突变表型的几个基因很可能与人类对应，这可能会导致相应的异常情况和出生缺陷。为了研究这些发育过程的生物学基础，无论是单个的，还是全部或全部的，这些基因被分子“克隆”是一个必要条件。虽然到目前为止已经克隆了几个这样的斑马鱼基因，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，特别是在大插入克隆试剂方面。许多发育生物学家仍然对重新启动这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力相当大，生物学家从解决与发育生物学密切相关的关键和有意义的问题到更深入地理解发育生物学的距离更远。正如跨NIH斑马鱼协调委员会所建议的那样，更好的基因组基础设施以促进突变基因的识别是社区的核心任务。因此，这项建议的首要目标是产生和维护斑马鱼界容易获得的大插入基因组资源，以便更快地克隆与发育相关的基因。这将通过以下方式实现：从近交系斑马鱼构建改良的BAC和PAC文库，生成这些文库的池以及现有的大插入库，向社区分发基因组试剂(以及适当的用户友好协议)，并作为基因组资源、信息和专业知识的中央储存库。与这些资源有关的数据将存储在ZFIN中，并将为其条目开发适当的脚本。传播实物和虚拟资源将鼓励社区内多层次、方案间的合作。此外，由此产生的文库将成为未来斑马鱼基因组大规模物理测绘和测序以及功能基因组学实验的不可或缺的组成部分。

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.