CLONING IMPRINTED GENES FROM THE MOUSE

基因组印记是指母体基因组中 和一个基因的父系等位基因。 各种各样的实验方法， 包括麦格拉思的经典核移植研究， 索尔特，帮助建立了适当的表达， 印记基因对哺乳动物的发育至关重要。 长期 这项研究的目的是使用小鼠模型来识别新的 印迹基因，并检查这些基因的功能， 和人类发展。 我们之前设计了一种新的育种方法 利用携带罗伯逊氏病的小鼠品系的策略 染色体产生的胚胎是单亲二体（UPD）， 7号染色体;换句话说，这条染色体的两个拷贝都被 来自母亲或父亲。 我们建议继续 通过对本试验的系统分析，划分了试验目标， 分为四个部分。 第一，新的印记基因将被识别， 使用比较RNA分析的UPD 7小鼠胚胎（差异 显示）。 第二，将得到的cDNA进行定位， 对7号染色体上的相应基因进行测序和评估， 等位基因特异性表达。 三、时空表达 将通过原位杂交检测所鉴定基因的模式 使用分阶段和切片的小鼠胚胎。第四，候选人资格 人类对应的印记小鼠基因作为疾病基因将是 评估。 预计这些研究不仅将提供 对哺乳动物中印记的性质和作用的新见解 同时，也将使我们更加了解 印记在人类疾病中扮演的角色， 综合征（PWS）、Angelman综合征（AS）、Beckwith-Wiedemann综合征 (BWS)和非细胞遗传学定义的微缺失综合征和UPD 方面的影响.

Genomic imprinting is defined as the unequal expression of the maternal and paternal alleles of a gene. A variety of experimental approaches, including the classical nuclear transplantation studies of McGrath and Solter, have helped to establish that appropriate expression of imprinted genes is critical to mammalian development. The long term goals of this research are to use a mouse model to identify novel imprinted genes and to examine the functions of these genes during mouse and human development. We have previously devised a novel breeding strategy which exploits mouse strains that carry Robertsonian chromosomes to generate embryos that are uniparental disomic (UPD) for chromosome 7; in other words, both copies of this chromosome have been derived from either the mother or the father. We propose to continue the systematic analysis of this cross, the experimental aims am divided into four Sections. One, novel imprinted genes will be identified from UPD 7 mouse embryos using comparative RNA analysis (differential display). Two, resulting cDNAs will be mapped to confirm the location of the corresponding genes on chromosome 7, sequenced and evaluated for allele-specific expression. Three, the temporal and spatial expression patterns of identified genes will be examined by in situ hybridization using staged and sectioned mouse embryos. Four, the candidacy of the human counterparts of imprinted mouse genes as disease genes will be evaluated. It is anticipated that these studies will not only provide novel insight into the nature and role(s) of imprinting in mammalian development but will also increase our understanding of the important role that imprinting plays in human diseases such as Prader-Willi syndrome (PWS), Angelman syndrome (AS), Beckwith-Wiedemann syndrome (BWS) and non-cytogenetically defined microdeletion syndromes and UPD effects.