Developing the Feline Radiation Hybrid Map

开发猫科动物辐射混合图

• 批准号: 6950950
• 负责人: STEPHEN J O'BRIEN
• 金额: --
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6950950
• 关键词: animal genetic material tag; biotechnology; cats; gene rearrangement; genetic mapping; genetic markers; genetic techniques; genome; nucleic acid sequence; polymerase chain reaction; sequence tagged sites; species difference; technology /technique development

Development of genomic resources in the domestic cat is key to application of this species as a model for heritable and infectious diseases, and for comparative genome analysis. Previous versions of the cat gene map, based on somatic cell hybrid analysis, revealed considerable conservation of synteny with the human genome. However, these maps provided no knowledge of gene order and genome rearrangement between the two species, information which is critical to applying the comparative candidate approach to positional cloning in gene-poor model systems. Radiation hybrid (RH) mapping has emerged as the most powerful tool for constructing moderate- to high-density gene maps in vertebrates by obviating the need to identify interspecific polymorphisms. A domestic cat RH panel has been developed and serves as the backbone for feline gene mapping, while also serving to cross-reference microsatellite positions on the current domestic cat-Asian leopard cat interspecies linkage map. We have completed a second generation 1150-locus map integrating 550 coding loci (expressed sequence tags, genes, and comparative anchor tagged sequences [CATS] markers) with 600 microsatellites selected from the current linkage map. By comparison to the genetic map, the 1150-marker RH map has a mean density of 1 genetic marker/5 cM, now useful for comparative candidate positional clone analysis of the over 200 described feline hereditary phenotypes. We are currently increasing the type I marker density to nearly 1000 loci through additional expressed sequence tag (EST) development and BAC-end sequencing-based marker production, focusing primarily on gaps in human-cat synteny. Presently we are also increasing microsatellite coverage at least five-fold with an ultimate goal of 1000-1500 microsatellites on the map. Further refinement of rearrangement breakpoints and conserved segment boundaries will enhance cross-reference to other mammalian species' genomes for use in diverse studies of genetics and genome evolution.

家猫基因组资源的开发是该物种作为遗传性和传染性疾病模型以及比较基因组分析的关键。先前版本的猫基因图谱，基于体细胞杂交分析，揭示了与人类基因组的同线性相当保守。然而，这些地图提供了两个物种之间的基因顺序和基因组重排的知识，这是关键的应用比较候选人的方法定位克隆在基因贫乏的模型系统的信息。辐射杂交（RH）作图已成为最有力的工具，建立中到高密度的脊椎动物基因图谱，避免了需要确定种间多态性。一个家猫RH面板已经开发出来，并作为猫基因定位的骨干，同时也用于交叉引用微卫星位置上目前的家猫，亚洲豹猫种间连锁图。我们已经完成了第二代1150个位点的地图整合550个编码位点（表达序列标签，基因，和比较锚标记序列[CATS]标记）与600个微卫星从目前的连锁图谱选择。与遗传图谱相比，1150标记RH图谱的平均密度为1个遗传标记/5 cM，现在可用于200多种描述的猫科动物遗传表型的比较候选位置克隆分析。我们目前正在通过额外的表达序列标签（EST）开发和基于BAC末端测序的标记生产，将I型标记密度增加到近1000个位点，主要集中在人-猫同线性的缺口上。目前，我们还将微型卫星的覆盖范围扩大至少五倍，最终目标是在地图上增加1 000至1 500颗微型卫星。重排断裂点和保守片段边界的进一步细化将增强与其他哺乳动物物种基因组的交叉引用，用于遗传学和基因组进化的各种研究。