GENOMIC RESOURCE DEVELOPMENT IN THE LABORATORY OPOSSUM

描述（改编自申请人的摘要）：灰色短尾 opossum，M.已被确立为模式生物， 对相关的广泛主题进行比较生物医学研究 对人类发育生理和疾病易感性的影响。 然而，缺乏 关于基本遗传特征的基本信息限制了其 潜在的调查涉及正常的遗传调节 发育和生理过程，以及遗传因素的影响 与健康相关的生理特征的变化。 为了缓解这种情况，M。南极洲的基因组将是 构建了可用于检测、定位并最终分离和克隆的 影响正常和异常表型变异的基因。 整体 目的是构建200个或更多多态性标记的连锁图 基因座，使用物理作图研究，以锚连锁图上的 物理基因组，并使用地图来确定一个或多个 影响饮食胆固醇和脂肪的可变反应的基因。 具体目标是：1）扩大匿名性和功能性 基因标记在现有的，稀疏填充的连锁图谱，通过继续 利用现有的回交作图板进行连锁研究; 2）完成5 cM连锁图，并利用新的回交作图法进行作图， 旨在最大限度地利用机会， 功能基因位点的多态性; 3）开发一个物理图谱，显示 已知的连锁群在M上的位置和方向。domestica 染色体;和4）检测和映射一个或多个基因负责 饮食诱导高胆固醇血症反应性的高度遗传变异 发生在这个物种。 该连锁图由多态性遗传标记位点组成， 匿名微卫星位点和功能基因，将开发 从基因型分析确定的基因间重组率分析 来自两个大型回交家族的后代 荧光原位 杂交（FISH）将被用来定位的物理位置， 已知的染色体连锁群。 参数LOD-评分关联 分析和谱系方差分量方法将被用来寻求 连锁图上的标记基因与影响 血浆脂蛋白表型在家庭小组已受到 饮食挑战方案。

DESCRIPTION (Adapted from the applicant's abstract): The gray-short-tailed opossum, M. domestica, has been established as a model organism for comparative biomedical research on a broad range of topics that are relevant to human development, physiology, and disease susceptibility. However, a lack of basic information on fundamental genetic characteristics limits its potential for inquiries involving the genetic regulation of normal developmental and physiologic processes, and the influences of genetic variation on health-related physiologic characteristics. To mitigate this situation, a map of the M. domestica genome will be constructed that can be used to detect, map, and ultimately isolate and clone genes that influence normal and abnormal phenotypic variation. The overall objectives are to construct a linkage map of 200 or more polymorphic marker loci, to use physical mapping studies to anchor the linkage map on the physical genome, and to use the map to determine the locations of one or more genes that influence variable responsiveness of dietary cholesterol and fat. The Specific Aims are to: 1) expand the number of anonymous and functional gene markers on the existing, sparsely-filled linkage map through continuing linkage studies utilizing an existing backcross mapping panel; 2) complete a 5 cM linkage map by creating, and utilizing for mapping, a new backcross mapping panel designed to maximize opportunities for the detection and mapping of polymorphisms at functional gene loci; 3) develop a physical map showing the locations and orientations of known linkage groups on M. domestica chromosomes; and 4) detect and map one or more of the genes responsible for highly heritable variation in diet-induced hypercholesterolemic responsiveness that occurs in this species. The linkage map, comprised of polymorphic genetic marker loci including anonymous micosatellite loci and functional genes, will be developed by analysis of intergenic recombination rates determined from genotype analyses of offspring from two large backcross family panels. Fluorescence in situ hybridization (FISH) will be used to locate the physical positions of the known linkage groups on individual chromosomes. Parametric LOD-score linkage analyses and pedigree based variance components methods will be used to seek linkage between marker genes on the linkage map and genes that influence plasma lipoprotein phenotypes in a family panel that has been subjected to a dietary challenge regimen.