CONSTRUCTION OF A GENETIC LINKAGE MAP OF ZEBRAFISH

脊椎动物胚胎学家和发育生物学家正在成为 对斑马鱼Brachydanio这种新的模型系统越来越感兴趣 再来一次。对斑马鱼的兴趣源于它的价值，既是一种 遗传和胚胎学有机体。这是遗传学的理想选择，因为 动物体型小，耐寒，容易大量饲养，因为： 雌鸟每次产卵100-200粒，每次产卵1-2次 每周，诱变和转基因技术一直很好- 已定义。对于胚胎学研究，他们的优势是受精 是外部的，胚胎几乎是透明的，所以迁徙 细胞和器官发生的最早阶段可以直接 观察到的。许多变异鱼在使用化学物质后被分离出来， 辐射诱变或插入诱变，以及至少三次诱变 目前，屏幕正在进行中，目标是在 接下来的几年。斑马鱼是唯一有这些的脊椎动物 优势。 具体来说，我们建议：L)构建第一个完整的基因 斑马鱼的连锁图谱。我们的目标是拥有一张至少有10分的地图 厘米分辨率。2)开发一个参考十字(1000条斑马鱼)，并绘制地图 克罗斯。该参考交叉对于启动精细结构是必不可少的 测绘，并促进位置克隆。参考十字将为 向整个斑马鱼社区提供。3)建立一种手段 资源分配，确保标记信息可供访问 调查人员，从参比十字中很容易获得DNA。 地图将提供什么，现在是投资建设 斑马鱼的遗传连锁图谱？一张地图将允许1)共时 保护地图；2)允许完全整合单一地图，以及 斑马鱼群落发现的突变的参考交叉；以及 3)为基因鉴定和定位克隆奠定基础。 斑马鱼养殖场正在迅速扩大，有数百只斑马鱼 研究人员目前正在研究斑马鱼和特定的斑马鱼突变。 如果地图建设被推迟，不仅资金将花费几个月 实验室制作特定地区的基本独立地图，但 还必须分配资金，以整合各种“地方” 地图。出于这些原因，经过讨论，最大的斑马鱼 设施决定重点构建遗传连锁图谱，使用 MGH的简单序列长度多态(SSLP)。就是为了这个 申请资金的原因。

Vertebrate embryologists and developmental biologists are becoming increasingly interested in a new model system, the zebrafish, Brachydanio rerio. The interest in zebrafish is derived from its value as both a genetic and embryological organism. It is ideal for genetics because the animals are small, hardy and easy to raise in large numbers because: the females lay 100-200 eggs each spawning, which can occur 1-2 times per week, and techniques for mutagenesis and transgenesis have been well- defined. For embryological studies, their advantage is that fertilization is external, and the embryos are nearly transparent, so that the migration of cells and the earliest stages of organogenesis can be directly observed. Many mutant fish have been isolated after using chemical, radiation, or insertional mutagenesis, and at least three mutagenesis screens are currently underway with the goal of reaching saturation within the next few years. The zebrafish is the only vertebrate with these advantages. Specifically we propose to: l) Construct the first complete genetic linkage map of the zebrafish. Our goal is to have a map with at least a 10 cM resolution. 2) Develop a reference cross (1000 zebrafish), and map this cross. This reference cross is essential for initiating fine structure mapping, and facilitating positional cloning. The reference cross will be made available to the entire zebrafish community. 3) Establish a means of resource distribution, insuring marker information is accessible to investigators, and that DNA from the reference cross is easily obtainable. What would the map provide and is this the time to invest in building a genetic linkage map in zebrafish? A map would 1) allow synteny conservation mapping; 2) allow complete integration of a single map, and reference cross for mutations discovered by the zebrafish community; and 3) build the foundation for gene identification and positional cloning. The zebrafish field is rapidly expanding, and there are hundreds of investigators now working on zebrafish and specific zebrafish mutations. If map construction is delayed, not only will funds be expended by several laboratories to make rudimentary independent maps of specific regions, but funds will also have to be allocated to integrate the various "local" maps. For these reasons, and after discussion, the largest zebrafish facilities decided to focus on construction of genetic linkage map using simple sequence length polymorphisms (SSLPs) at the MGH. It is for this reason funds are requested.