Synthesis, Organization and Function of Ribonucleic Acid

9418632佩里这项研究的总体目标是阐明控制真核细胞中基因表达的分子机制。这些研究旨在了解一种有趣的DNA结合蛋白CHD1在决定高阶染色质结构和调节基因活性方面的作用。CHD1具有某些结构特征(一个染色质结构域和一个ATPase/解旋酶结构域)，这与参与转录调控的其他染色质相关蛋白有关。将使用广泛的遗传和生化技术，包括在各种培养细胞中的DNA转基因和显微注射实验，免疫荧光分析，柱层析纯化蛋白质，基因克隆和测序，DNA、RNA和免疫印迹，引物延伸和核苷酸保护分析，电泳迁移率变化和DNA足迹分析，以及在细菌中生产野生型和突变蛋白。这项研究将在哺乳动物细胞、果蝇和酵母中进行。这些研究的结果应该有助于我们对基因调控的认识，因为这些因素决定了基因与转录装置的可及性，并有助于介导细胞周期依赖的染色质凝聚变化。尽管在过去的几年里积累了大量关于基因调控的令人印象深刻的事实，但这里所解决的一般问题在很大程度上仍然没有得到解决。这项研究的总体目标是阐明控制真核细胞中基因表达的分子机制。这些研究旨在了解一种有趣的DNA结合蛋白CHD1在决定高阶染色质结构和调节基因活性方面的作用，CHD1具有某些结构特征，这些结构特征与其他与转录调控有关的染色质相关蛋白有关。将使用广泛的遗传和生化研究技术，并将对哺乳动物细胞、果蝇和酵母进行研究。尽管在过去的几年里积累了大量关于基因调控的令人印象深刻的事实，但这里所解决的一般问题在很大程度上仍然没有得到解决。这些研究的结果通过定义决定基因与转录装置的可及性的因素，以及有助于介导细胞周期依赖的染色质结构变化的因素，应该大大有助于我们对基因调控的了解。*？？

9418632 Perry The overall goal of this research is to elucidate molecular mechanisms that govern gene expression in eukaryotic cells. The proposed studies are aimed at understanding the role of an interesting DNA-binding protein, CHD1, in the determination of higher-order chromatin structure and the regulation of gene activity. CHD1 possesses certain structural features (a chromodomain and an ATPase/helicase domain), which relate it to other groups of chromatin-associated proteins that have been implicated in transcriptional regulation. A broad spectrum of genetic and biochemical techniques will be used, including DNA transfection and microinjection experiments in a variety of cultured cells, immunofluorescence analyses, protein purification by column chromatography, gene cloning and sequencing, DNA-, RNA- and immuno-blotting, primer extension and nucleaseprotection assays, electrophoretic-mobility shift and DNA footprint analyses, and the production of wild-type and mutant proteins in bacteria. The studies will be carried out with mammalian cells, Drosophila and yeast. The results of these studies should contribute substantially to our knowledge of gene regulation by defining factors that determine the accessibility of genes to the transcriptional apparatus and that help mediate cell-cycle dependent changes in chromatin condensation. Although an impressive body of facts pertaining to gene regulation has accumulated over the past several years, the general problems being addressed here remain largely unsolved. %%% The overall goal of this research is to elucidate molecular mechanisms that govern gene expression in eukaryotic cells. The proposed studies are aimed at understanding the role of an interesting DNA-binding protein, CHD1, in the determination of higher-order chromatin structure and the regulation of gene activity, CHD1 possesses certain structural features which relate it to other groups of chromatin-associated proteins that have been implicated in transcriptional regulation. A broad spectrum of genetic and biochemical investigative techniques will be used and studies will be carried out with mammalian cells, Drosophila and yeast. Although an impressive body of facts pertaining to gene regulation has accumulated over the past several years, the general problems being addressed here remain largely unsolved. The results of these studies should contribute substantially to our knowledge of gene regulation by defining factors that determine the accessibility of genes to the transcriptional apparatus and that help mediate cell-cycle dependent changes in chromatin structure. *** ??