Synthesis, Organization and Function of Ribonucleic Acid

9418632佩里这项研究的总体目标是阐明真核细胞中基因表达的分子机制。拟议的研究旨在了解一个有趣的DNA结合蛋白质，CHD 1，在确定高阶染色质结构和基因活性的调节中的作用。CHD 1具有某些结构特征（一个染色质结构域和一个ATP酶/解旋酶结构域），这使它与其他组的染色质相关蛋白质有关，这些蛋白质与转录调控有关。 将使用广泛的遗传和生物化学技术，包括在各种培养细胞中进行DNA转染和显微注射实验、免疫荧光分析、柱色谱法纯化蛋白质、基因克隆和测序、DNA、RNA和免疫印迹、引物延伸和核酸酶保护测定、细胞迁移率变化和DNA足迹分析，以及在细菌中产生野生型和突变蛋白。这些研究将用哺乳动物细胞、果蝇和酵母进行。 这些研究的结果应大大有助于我们的基因调控的知识，通过定义的因素，确定基因的转录装置的可及性，并帮助介导细胞周期依赖的染色质凝聚的变化。虽然在过去的几年里，已经积累了大量与基因调控有关的事实，但这里所讨论的一般问题在很大程度上仍然没有解决。 本研究的总体目标是阐明真核细胞中基因表达的分子机制。 拟议的研究旨在了解一个有趣的DNA结合蛋白质，CHD 1，在确定高阶染色质结构和基因活性的调节中的作用，CHD 1具有一定的结构特征，将其与其他组的染色质相关的蛋白质，这些蛋白质与转录调控有关。 将使用广泛的遗传和生物化学调查技术，并将对哺乳动物细胞、果蝇和酵母进行研究。 虽然在过去的几年里，已经积累了大量与基因调控有关的事实，但这里所讨论的一般问题在很大程度上仍然没有解决。 这些研究的结果应大大有助于我们的基因调控的知识，通过定义的因素，决定了基因的可及性的转录装置，并有助于介导细胞周期依赖的染色质结构的变化。 *** ??

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. *** ??