DELETION MAPPING OF THE ZEBRAFISH GENOME

描述(改编自申请人的摘要)：申请 建议确定细胞和分子成分对 对早期组织的分化和空间组织 脊椎动物胚胎。事实证明，其中一条途径提供了特别多的信息 为了阐明单个细胞的命运和整体的身体计划 在无脊椎动物身上确定的是对发育的分析 突变。最近对斑马鱼(Brachydanio Rerio)的研究 证明了这种脊椎动物具有许多 发育遗传学研究。这项研究的长期目标是 对符合以下条件的基因进行详细的遗传和分子表征 有助于早期发展，位于两个地区中的任何一个 由白化病和黄色基因座定义的基因组。这样做的目的是 应用是建立一系列重叠的缺失突变 在白化病或黄色基因座附近。可遗传缺陷突变将是 对发育中的斑马鱼胚胎进行伽马辐照后的诱导。 总共500条诱变的鱼将与窝藏在 白化或黄色的隐性、有活力的色素沉着突变 识别新诱导的非互补性突变的传播。 大约1%的诱变鱼预计会携带 每个预先指定的基因座都有一个新的突变。每一个被诱导的 基因突变将被定性为基因突变。这些功能被 突变将通过检查表型进行分析 纯合突变胚胎。两者之间的配对互补检验 诱导的突变将：1)识别缺失突变 影响多个基因功能；2)帮助确定 每一系列隐性致死对不同基因功能的影响 突变；以及3)允许为白化病构建缺陷图 和基因组的黄色区域。为了直接证明其中一些 大鼠诱导的突变均为缺失，纯合子突变将被 分析50个特定DNA序列基因座中的任何一个的丢失。建议数 项目代表了一种有效的恢复 斑马鱼中的发育突变。缺失突变将 代表着向完整的基因和分子迈出的第一步 白化病和黄化病附近早期调控基因的特征 发展。与这些相关的发育缺陷的分析 突变将揭示细胞和分子的基本功能 脊椎动物的早期胚胎发育。

DESCRIPTION (Adapted from the applicant's abstract): The application proposes to identify cellular and molecular components that contribute to the differentiation and spatial organization of tissues in the early vertebrate embryo. One avenue that has proven particularly informative for elucidating how individual cell fates and overall body plan are determined in invertebrates has been the analysis of developmental mutations. Recent studies with the zebrafish (Brachydanio rerio) have proven that this vertebrate has many of the attributes required for developmental genetic studies. The longterm goal of the study is to perform a detailed genetic and molecular characterization of genes that contribute to early development and that lie in either of two regions of the genome defined by the albino and yellow loci. The goal of this application is to establish a series of overlapping deletion mutations near the albino or yellow loci. Heritable deficiency mutations will be induced following gamma irradiation of developing zebrafish embryos. A total of 500 mutagenized fish will be mated with individuals harboring recessive, viable, pigmentation mutations at albino or yellow to identify the transmission of newly induced, non-complementing mutations. Approximately 1 percent of the mutagenized fish are expected to harbor a new mutation at each pre-specified locus. Each of the induced mutations will be characterized genetically. The functions perturbed by the mutations will be analyzed by examination of the phenotypes of homozygous mutant embryos. Pairwise complementation tests between each of the induced mutations will: 1) identify deletion mutations that affect several gene functions; 2) help determine the minimal number of distinct gene functions affected by each series of recessive lethal mutations; and 3) permit construction of deficiency maps for the albino and yellow regions of the genome. To demonstrate directly that some of the rat-induced mutations are deletions, homozygous mutants will be analyzed for loss of any of 50 specific DNA sequence loci. The proposed project represents an efficient approach to the recovery of developmental mutants in the zebrafish.The deletion mutations will represent a first step toward complete genetic and molecular characterization of the genes near albino and yellow that regulate early development. Analysis of the developmental defects associated with these mutations will reveal cellular and molecular functions that are essential to early vertebrate embryogenesis.