Hagiwara Y, Hirai M, Nishiyama K, Kanazawa I, Ueda T, Sakaki Y, Ito T
Human Genome Center, Institute of Medical Science, University of Tokyo, 4-6-1 Shirokane-dai, Minato-ku, Tokyo 108, Japan.
Proc Natl Acad Sci U S A. 1997 Aug 19;94(17):9249-54. doi: 10.1073/pnas.94.17.9249.
A systematic screen termed the allelic message display (AMD) was developed for the hunting of imprinted genes. In AMD, differential display PCR is adopted to image allelic expression status of multiple polymorphic transcripts in two parental mouse strains, reciprocal F1 hybrids and pooled backcross progenies. From the displayed patterns, paternally and maternally expressed transcripts can be unequivocally identified. The effectiveness of AMD screening was clearly demonstrated by the identification of a paternally expressed gene Impact on mouse chromosome 18, the predicted product of which belongs to the YCR59c/yigZ hypothetical protein family composed of yeast and bacterial proteins with currently unknown function. In contrast with previous screening methods necessitating positional cloning efforts or generation of parthenogenetic embryos, this approach requires nothing particular but appropriately crossed mice and can be readily applied to any tissues at various developmental stages. Hence, AMD would considerably accelerate the identification of imprinted genes playing pivotal roles in mammalian development and the pathogenesis of various diseases.
为寻找印记基因,开发了一种名为等位基因信息显示(AMD)的系统筛选方法。在AMD中,采用差异显示PCR来成像两个亲本小鼠品系、正反交F1杂种和混合回交后代中多个多态性转录本的等位基因表达状态。从显示的模式中,可以明确鉴定出父本和母本表达的转录本。通过鉴定小鼠18号染色体上一个父本表达的基因Impact,清楚地证明了AMD筛选的有效性,其预测产物属于由酵母和细菌蛋白质组成的YCR59c/yigZ假设蛋白家族,目前其功能未知。与以往需要进行定位克隆或孤雌生殖胚胎的筛选方法不同,这种方法只需要适当杂交的小鼠,不需要任何特殊条件,并且可以很容易地应用于不同发育阶段的任何组织。因此,AMD将大大加速对在哺乳动物发育和各种疾病发病机制中起关键作用的印记基因的鉴定。