CLONING IMPRINTED GENES FROM THE MOUSE

基因组印记被定义为母体的不平等表达 和父亲的等位基因。各种实验方法， 包括McGrath的经典核移植研究和 Solter，帮助建立了适当表达 印记基因对哺乳动物的发育至关重要。从长远来看 这项研究的目标是使用小鼠模型来识别新奇的 印迹基因并检测这些基因在小鼠体内的功能 和人的发展。我们之前设计了一种新的繁殖方式 利用携带罗伯逊病毒的小鼠品系的策略 染色体产生的胚胎是单亲二体(UPD) 7号染色体；换句话说，这条染色体的两个副本都是 从母亲或父亲那里得到的。我们建议继续 对该交叉的系统分析，将实验目的划分为 分为四个部分。第一，新的印记基因将从 使用比较RNA分析的UPD 7小鼠胚胎(差异 显示)。第二，生成的cDNA将被映射以确认位置 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 已鉴定基因的模式将通过原位杂交进行检查 使用分段和分段的小鼠胚胎。四、候选人资格 人类印记的小鼠基因作为疾病基因将被 已评估。预计这些研究不仅将提供 对哺乳动物印记的性质和作用的新见解(S) 发展，但也将增加我们对 印记在诸如Prader-Willi等人类疾病中的作用 综合征(PWS)、安杰曼综合征(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 效果。

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.