CLONING AND CHARACTERIZING NEW IMPRINTED GENES

描述：(改编自PI的摘要)本项目致力于 基因组印迹的功能及其潜在的调控机制。 该项目的目标包括：1)识别额外的鼠标 印记基因，以便建立比较工作的基础 人类基因组。使用限制性地标基因组扫描(RLGS)方法， 确定了显示等位基因特异性甲基化的候选基因座；a 导致新印记基因克隆的印记基因的共同特征 基因。现在计划再克隆11个候选基因座 大片段相邻基因的初步鉴定及鉴定 其中一人的基因组克隆。2)详细描述了新的 鉴定小鼠9号染色体上的印迹区域，包括父本 表达了鸟核苷酸交换因子(Grf1)基因。Grf1是鼠标 与酵母CDC25基因同源，CDC25基因作为RAS激活剂，因此 在细胞周期调控中起着重要作用。白纹伊蚊的基因组结构 我们将分析一组基因组克隆中500kbp重叠群内的Grf1基因。 这些实验将包括外显子-内含子的特征 Grf1的结构、额外等位基因特异性甲基化的鉴定 位点和外显子捕获实验以识别新的编码序列。3) 不同选择性剪接基因表达的详细研究 Grf1成绩单的形式，包括识别和分析 最大的父系表达的Grf1转录本的启动子。

DESCRIPTION: (adapted from the PI's abstract) This project addresses the function of genomic imprinting and the underlying regulatory mechanisms. The goals of the project include: 1) Identification of additional mouse imprinted genes in order to establish the basis for comparative work in the human genome. Using the Restriction Landmark Genome Scanning (RLGS) method, candidate loci were identified which show allele-specific methylation; a common feature of imprinted genes that led to the cloning of novel imprinted genes. It is now planned to clone 11 additional candidate loci for preliminary characterization and to identify the adjacent gene in large genomic clones for one of them. 2) A detailed characterization of the newly identified imprinted region on mouse chromosome 9 including the paternally expressed Grf1 (guanine nucleotide exchange factor) gene. Grf1 is the mouse homologue to the yeast CDC25 gene which acts as a Ras activator and thus plays an important role in cell cycle regulation. The genomic structure of the Grf1 gene within a 500 kbp contig of genomic clones will be analyzed. These experiments will include the characterization of the exon-intron structure of Grf1, identification of additional allele-specific methylation sites and exon-trapping experiments to identify novel coding sequences. 3) A detailed study on the expression of the different alternatively spliced forms of Grf1 transcripts, including the identification and analysis of the promoter for the largest, paternally expressed Grf1 transcript.