CONSTRUCTION OF A GENETIC LINKAGE MAP OF ZEBRAFISH

此拨款申请是为了响应基因组RFA而提交的 资源的斑马鱼，要求申请资金， 在斑马鱼中构建0.3cM的图谱。 我们响应RFA的目标 是生成斑马鱼Danio的密集微卫星连锁图 rerio，一个将对克隆基因的容易性产生巨大影响的项目， 以一种方便的方式，以一种方便的方式， 科学家们使用这种生物。 超过600个发育重要基因已被确定， 斑马鱼通过化学诱变筛选。 我们学院是一个专业 这种筛选和克隆被破坏的基因，特别是 与器官发育有关的。这些变异说明 重要的人类疾病，包括心力衰竭和心律失常， 癌症，造血缺陷，视网膜变性，多囊 肾病和神经退化 这些基因的分离将 需要一个密集和强大的遗传图谱，由可靠的，易于 使用可用于多种鱼类杂交的标记。 基于简单序列长度多态性的微卫星标记 （SSLPs）在整个基因组中的重复现在是标准的开始。 定位克隆和锚定其他地图在大多数遗传 有机体 它们在整个基因组中分布丰富; 它们在远交品系之间具有高度多态性;与RAPD不同， 它们的高共显性发生率使它们特别有用， 二倍体和非近交系;它们是单位点扩增 产品，使其成为锚定辐射的重要工具 基于杂交（RH）的表达序列标签（EST）映射到遗传图谱， 这也是RFA的另一个目标。 在我们的地图项目的最初几年，我们开发了一个低- 密度完整的地图，有足够的标记（600-700），以覆盖所有25个 染色体的平均间隔为5厘摩（cM）。 许多 世界各地的研究人员利用我们目前的低密度 微卫星图谱启动重要基因的定位克隆， 但同意我们作为一个领域，必须增加 我们地图上的微卫星标记，并使它们迅速可用。 我们 实验室，建立在取得的经验，同时使低 密度图，已经开发了一种简化的方法来做到这一点。

This grant application is submitted in response to the RFA for Genomic Resources for the Zebrafish, requesting applications for funds to construct a 0.3 cM map in zebrafish. Our goal in responding to this RFA is to generate a dense microsatellite linkage map of the zebrafish Danio rerio, one that will have a dramatic effect on the ease of cloning genes by position, and to do so in a manner of ready utility for the entire community of scientists using this organism. More than 600 genes of developmental importance have been defined in zebrafish by chemical mutagenesis screens. Our institute is one major site for such screens and the cloning of the disrupted genes, especially those which are relevant to organ development. The mutations speak to important human diseases, including heart failure and arrhythmias, cancer, hematopoietic deficiency, retinal degeneration, polycystic kidney disease, and neural degeneration. Isolation of these genes will require a dense and robust genetic map, composed of reliable, easy-to- use markers that can be used in crosses between many strains of fish. Microsatellite markers based on simple sequence length polymorphisms (SSLPs) repeats throughout the genome are now the standard starting point for positional cloning and anchoring of other maps in most genetic organisms. They are abundant and well distributed through the genome; they are highly polymorphic et between outbred strains; unlike RAPDs, their high incidence of codominance makes them especially useful in diploids and non-inbred strains; and they are single locus amplification products, making them an essential tool for the anchoring of a radiation hybrid (RH)-based expressed sequence tag (EST) map to the genetic map, which is the other component goal of this RFA. During the initial years of our mapping project, we developed a low- density complete map with sufficient markers (600-700) to cover all 25 chromosomes at an average interval of 5 centiMorgans (cM). Many investigators around the world have used our current low-density microsatellite map to initiate positional cloning of important genes, but agree that we, as a field, must increase the number of microsatellite markers on our map and make them rapidly available. Our laboratory, building on the experience gained while making the low density map, has developed a streamlined method to do so.