SFM VISUALIZATION OF SINGLE CHROMATIN FIBER DYNAMICS

描述：(应聘者？S描述) 该项目的主要目标是在缓冲液中单个核小体或纤维水平上进行生化实验，如扫描力显微镜所示。 实验计划大致可分为三个部分： (1)在缓冲液中对单个纤维进行可逆压实，然后进行SFM。同一纤维在低盐、中盐和高盐中的图像将被用于生成纤维结构的模型。为此，将创造一种新的、相位良好的光纤。 (2)当伸长的RNA聚合酶遇到核小体时会发生什么？实验旨在询问当单个RNA聚合酶实时遇到缓冲区中的单个核小体时会发生什么。 (3)小鼠乳腺肿瘤病毒(MMTV)长末端重复序列中的核小体如何与转录因子相互作用？特定的核小体是否发生了移位，或者它们的结构是否发生了改变，以允许转录因子结合？

DESCRIPTION: (Applicant?s Description) The major goals of this project are to perform biochemical experiments at the single nucleosome or fiber level in buffer as visualized by scanning force microscopy. Experimental plans can be roughly separated into three parts: (1) Reversible compaction of individual fibers followed in buffer by SFM. Images of the same fiber in low, moderate and high salts will be used to generate models of fiber structure. A new, well-phased fiber will be created for this purpose. (2) What occurs when an elongating RNA polymerase encounters a nucleosome? Experiments are designed to ask what happens when a single RNA polymerase encounters a single nucleosome in buffer in real time. (3) How do nucleosomes in the Mouse Mammary Tumor Virus (MMTV) Long Terminal Repeat (LTR) interact with transcription factors? Are specific nucleosomes displaced or are their structures altered to allow transcription factor binding?