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- W158943628 abstract "Here, we describe methods to prepare a mammalian expression plasmid encoding EGFP fused to the amino-terminus of human DNA topoisomerase IIα (Topo IIα) for use in studying the dynamics of Topo IIα in living cells. In previous studies, this plasmid was transfected into LLC-Pk cells, a porcine epithelial cell line that remains relatively flat during mitosis. After selection for stable integration, cells were cloned by serial dilution in microwells and used to grow a stable cell line expressing EGFP-Topo IIα; this cell line was termed LPk-GT2. Using photobleaching methods with conventional and patterned photobleaching, LPk-GT2 cells were used to demonstrate the rapid dynamics of Topo IIα exchange in both interphase nuclei and mitotic chromosomes. These rapid dynamics are dependent on enzyme activity since ICRF159, a catalytic inhibitor of Topo IIα, slows dynamics significantly. The methods utilized in these studies are described herein." @default.
- W158943628 created "2016-06-24" @default.
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- W158943628 date "2009-01-01" @default.
- W158943628 modified "2023-09-26" @default.
- W158943628 title "The Dynamics of DNA Topoisomerase IIα in Living Cells" @default.
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- W158943628 doi "https://doi.org/10.1007/978-1-60761-340-4_18" @default.
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