A Phosphorimager for Analyses of the Molecular Genetics of Signal Transduction, DNA Replication and Gene Expression

一个多用户团体建议购买一台磷成像仪，用于对信号转导、DNA复制和基因表达的分子遗传学分析中的放射性和化学发光物种进行快速、灵敏和准确的检测和定量。具体项目包括：分析真核细胞中DNA复制的启动机制和这一过程是如何调控的；受体酪氨酸激酶激活RAS蛋白的机制和确定激活的RAS蛋白的下游靶标；通过RNA聚合酶II剖析调节真核细胞基因转录的蛋白质-蛋白质相互作用；测试大RNA分子结构的模型和确定RNA结构可以影响基因表达的方式，以及探索参与细胞生长和发育的信号转导途径。将利用磷成像仪的具体应用包括蛋白质的体内放射性标记；蛋白质半衰期的测定；蛋白质翻译后修饰、亚细胞定位和寡聚体状态的分析；蛋白质-蛋白质、蛋白质-核酸和核酸内相互作用的分析；转录调控元件的分析；体内和体外DNA复制产品的分析。这些研究的结果将大大扩展我们对基本细胞过程的理解。

A multi-user group proposes to purchase a phosphorimager for the rapid, sensitive, and accurate detection and quantitation of radioactive and chemiluminescent species in analyses of the molecular genetics of signal transduction, DNA replication, and gene expression. Specific projects include analysis of the mechanism by which DNA replication is initiated in eukaryotic cells and how this process is regulated; the mechanism of activation of Ras proteins by receptor tyrosine kinases and the identification of downstream targets of activated Ras proteins; the dissection of protein-protein interactions that regulate transcription of genes in eukaryotic cells by RNA polymerase II; testing models for the structure of large RNA molecules and determination of the ways in which RNA structure can influence gene expression, and exploration of signal transduction pathways involved in cell growth and development. Specific applications that will utilize the phosphorimager include in vivo radiolabelling of proteins; determination of protein half-life; analysis of protein posttranslational modificatimns, subcellular localization and oligomeric states; analysis of protein-protein, protein-nucleic acid, and intranucleic acid interactions; analysis of transcriptional control elements; analysis of in vivo and in vitro DNA replication products. The results of these studies will significantly extend our understanding of basic cellular processes.