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DNA binding fluorescent proteins as singlemolecule probes

机译:DNA将荧光蛋白结合为单分子探针

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摘要

DNA binding fluorescent proteins are useful probes for a broad range of biological applications. Fluorescent protein (FP)-tagging allows DNA binding proteins expressed within a living cell to be directly visualised, in real-time, to study DNA binding patterns and dynamics. Moreover, FP-tagged DNA binding proteins (FP-DBP) have allowed the imaging of single proteins bound to large elongated DNA molecules with a fluorescence microscope. Although there are numerous DNA binding proteins, only a small portion of them have been exploited to construct FP-DBPs to study molecular motion in a cell or in vitro singlemolecule visualisation. Therefore, it would be informative to review FP-DBP for further development. Here, we summarise the design of FP-DBPs and their brightness, photostability, pK_a, maturation rate, and binding affinity (K_d) characteristics. Then, we review the applications of FP-DBP in cells to study chromosome dynamics, DNA replication, transcription factors, DNA damage, and repair. Finally, we focus on single DNA molecule visualisation using FP-DBP.
机译:DNA结合荧光蛋白是用于广泛的生物应用的有用探针。荧光蛋白(FP) - 可允许在活细胞内表达的DNA结合蛋白实时地直接可视化,以研究DNA结合模式和动力学。此外,FP标记的DNA结合蛋白(FP-DBP)允许用荧光显微镜与大细长DNA分子结合的单一蛋白质的成像。虽然存在许多DNA结合蛋白,但是它们的一小部分已被利用以构建FP-DBPS,以研究细胞或体外单点可视化中的分子运动。因此,审查FP-DBP以进一步发展,这将是丰富的。在这里,我们总结了FP-DBP的设计及其亮度,光稳定性,PK_A,成熟率和结合亲和力(K_D)特性。然后,我们审查FP-DBP在细胞中的应用,以研究染色体动力学,DNA复制,转录因子,DNA损伤和修复。最后,我们专注于使用FP-DBP的单一DNA分子可视化。

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