The Development of the Domestic cat, Felis catus, as an

家猫（Felis catus）的发展

• 批准号: 6558902
• 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/6558902
• 关键词: animal genetic material tag; animal population genetics; biochemical evolution; cats; genetic disorder; genetic markers; genetic models; genetic polymorphism; genetic susceptibility; genome; linkage mapping; model design /development; nucleic acid repetitive sequence; nucleic acid sequence; polymerase chain reaction; species difference

A goal of our research is to characterize genetic organization in the domestic cat and to develop genomic resources facilitating and establishing Felis catus as a useful animal model contributing to our understanding of human hereditary disease analogues, neoplasia, genetic factors associated with host response to infectious disease and mammalian genome evolution. In order to map and characterize genes associated with inherited pathologies in the domestic cat which mirror inherited human conditions we have recently completed a second generation linkage map of integrated microsatellite (Type II) and Type I (coding genes) in the domestic cat. The map was generated in a 134-member, multi-generation backcross pedigree between the domestic cat and the Asian leopard cat (Prionailurus bengalensis) and consists of 334 markers including 81 Type I loci and 253 Type II loci (246 autosomal and 7 X-linked microsatellites. The average intragroup inter-marker distance of 11 centimorgans (cM) was observed over the 18 feline autosomes and the X chromosome. In order to increase marker density for greater genetic linking mapping resolution in disease pedigrees, we have isolated an additional 700 dinucleotide repeat loci from a (dCA.dGT) enriched library. Primers for amplification have been designed and tested for both product performance and locus heterozygosity in a panel of outbred domestic cats. The loci are currently being genotyped in a cat x rodent radiation hybrid panel, recently used in construction of a Type I/Type II radiation hybrid map (see ZO1 BC 10265-05 LGD). A highly efficient approach is being utilized with amplifications performed in 384-well format and traditional gel-based assays have been replaced by a homogeneous 5'-nuclease assay involving a single common probe for all genetic markers. Following genotyping, the markers will be integrated with the current radiation hybrid map of the cat, including 421 Type I and 265 Type II markers. Microsatellites mapped at regular physical intervals will be genotyped and mapped in a large multi-generation domestic cat pedigree to generate a genetic linkage map with full genome coverage of Felis catus and of sufficient resolution for linkage mapping in disease pedigrees.

本研究的目的是描述家猫的遗传组织特征，开发基因组资源，促进并建立猫科动物作为有用的动物模型，有助于我们理解人类遗传病类似物、肿瘤、与宿主对传染病的反应相关的遗传因素以及哺乳动物基因组进化。为了定位和鉴定反映人类遗传条件的家猫遗传病理相关基因，我们最近完成了家猫整合微卫星(类型II)和类型I(编码基因)的第二代连锁图谱。 该图谱由334个标记组成，包括81个I型基因座和253个II型基因座(246个常染色体微卫星和7个X连锁微卫星)。在18个猫常染色体和X染色体上观察了11个厘米器官的平均群内标记间距离。为了在疾病家系中增加标记密度以获得更高的遗传连锁图谱分辨率，我们从(dCA.dGT)丰富的文库中额外分离了700个二核苷酸重复基因座。设计了用于扩增的引物，并在一组近交家猫身上对产品性能和基因座杂合性进行了测试。这些基因座目前正在CAT x啮齿动物辐射杂交仪表板上进行基因分型，最近用于构建I/II型辐射杂交图谱(见ZO1 BC 10265-05LGD)。正在利用一种高效的方法进行384孔格式的扩增，传统的基于凝胶的分析已被均相5‘-核酸酶分析所取代，该均相5’-核酸酶分析涉及所有遗传标记的单一公共探针。基因分型后，这些标记将与目前猫的辐射杂交图谱整合，包括421个I型和265个II型标记。以固定的物理间隔绘制的微卫星将在一个大型的多世代家猫家系中进行基因分型和定位，以生成一个完全覆盖猫科动物基因组并具有足够分辨率的遗传连锁图谱，用于疾病家系的连锁图谱绘制。