Molecular Cytogenetic Tools for the Zebrafish

斑马鱼的分子细胞遗传学工具

• 批准号: 6929094
• 负责人: CHARLES LEE
• 金额: $ 31.92万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-08-01 至 2009-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6929094
• 关键词: Internet; alternatives to animals in research; artificial chromosomes; centromere; comparative genomic hybridization; cytogenetics; fluorescent in situ hybridization; genetic mapping; genetic markers; karyotype; linkage mapping; method development; molecular biology information system; molecular probes; nucleic acid sequence; telomere; zebrafish

DESCRIPTION (provided by applicant): The zebrafish is an important model organism for studying vertebrate development, cell biology and cancer. Large-scale mapping and sequencing efforts currently rely on piecing together randomly characterized DNA fragments. Molecular cytogenetic methodologies adds a unique and complementary mapping approach by visualizing the presence of DNA sequences at specific chromosomal sites. Unfortunately, molecular cytogenetic probes for the zebrafish are lacking. Molecular cytogenetic studies will bridge the gap between the physical and genetic maps and expedite the genetic dissection of zebrafish mutant models of human diseases. This application proposes to develop a comprehensive set of molecular cytogenetic tools for the zebrafish by integrating cytogenetic landmarks into the zebrafish genome project. This will be accomplished first by standardizing the zebrafish karyotype and correlating the physical chromosomes with the genetic linkage maps using specific near-centromeric BAC probes. Five hundred BACs will be mapped by fluorescence in situ hybridization (FISH) to wild type metaphase chromosomes and anchored to existing maps and contigs. Information obtained will be posted on an updatable and interactive zebrafish cytogenetic website. The second objective will be to develop chromosome-specific FISH probes and an array CGH platform. BAC probes will be identified which map near the centromere and telomeres of each zebrafish chromosome. These probes will be used to characterize gamma-irradiated mutant lines thought to contain deletions or chromosomal translocations. Also, "paint probes" for each zebrafish chromosome will be produced and used to characterize interspecific chromosomal rearrangements and supernumerary marker chromosomes. Finally, a 3 Mb resolution BAC-based array CGH platform will be constructed. This tool will be used to identify genomic DNA gains and losses which could lead to the identification of novel oncogenes and tumor-suppresor genes that lead to tumorigenesis in many zebrafish mutants.

描述（由申请人提供）：斑马鱼是研究脊椎动物发育，细胞生物学和癌症的重要模型生物。当前的大规模映射和测序工作依赖于将随机表征的DNA片段拼凑在一起。分子细胞遗传学方法论通过可视化在特定染色体部位的DNA序列的存在，添加了一种独特的互补映射方法。不幸的是，缺乏斑马鱼的分子细胞遗传学探针。分子细胞遗传学研究将弥合物理图和遗传图之间的差距，并加快人类疾病的斑马鱼突变模型的遗传解剖。该应用建议通过将细胞遗传学的地标整合到斑马鱼基因组项目中，为斑马鱼开发一组全面的分子细胞遗传学工具。这将首先通过标准化斑马鱼核型，并使用特定的近中央胶粒BAC探针将物理染色体与遗传连锁图相关联。将通过荧光原位杂交（FISH）与野生型中期染色体绘制五百个BAC，并锚定在现有的地图和重叠群上。获得的信息将发布在可更新且互动的斑马鱼细胞遗传学网站上。第二个目标是开发特定于染色体的鱼类探针和阵列CGH平台。将鉴定出每个斑马鱼染色体的丝粒和端粒附近的哪个图。这些探针将用于表征被认为包含缺失或染色体易位的伽马射线突变系。另外，将生产每个斑马鱼染色体的“油漆探针”，并用于表征种间染色体重排和超努标记染色体。最后，将构建一个3 MB分辨率的基于BAC的阵列CGH平台。该工具将用于鉴定基因组DNA的增长和损失，这可能导致新的癌症基因和肿瘤供应基因，这些基因导致许多斑马鱼突变体的肿瘤发生。