Abstract
DNA supercoiling plays critical roles in several essential DNA metabolic pathways, such as replication, transcription and recombination. Typically plasmid DNA molecules are used to measure DNA supercoiling status inside bacterial cells. In this chapter, we describe how to isolate plasmid DNA molecules from E. coli cells and determine DNA supercoiling density by 1% agarose gel electrophoresis containing chloroquine using plasmid pACYC184 as an example.
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References
Champoux JJ (2001) DNA topoisomerases: structure, function, and mechanism. Annu Rev Biochem 70:369–413
Snoep JL, van der Weijden CC, Andersen HW, Westerhoff HV, Jensen PR (2002) DNA supercoiling in Escherichia coli is under tight and subtle homeostatic control, involving gene-expression and metabolic regulation of both topoisomerase I and DNA gyrase. Eur J Biochem 269:1662–1669
Wang JC (1996) DNA topoisomerases. Annu Rev Biochem 65:635–692
Zechiedrich EL, Khodursky AB, Bachellier S, Schneider R, Chen D, Lilley DM, Cozzarelli NR (2000) Roles of topoisomerases in maintaining steady-state DNA supercoiling in Escherichia coli. J Biol Chem 275:8103–8113
White JH, Bauer WR (1986) Calculation of the twist and the writhe for representative models of DNA. J Mol Biol 189:329–341
Cozzarelli NR, Boles TC, White JH (1990) A primer on the topology and geometry of DNA supercoiling. In: Cozzarelli NR, Wang JC (eds) DNA topology and its biological effects. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, pp 139–184
Bates AD, Maxwell A (2005) DNA topology, 2nd edn. Oxford University Press, Oxford
White JH, Cozzarelli NR, Bauer WR (1988) Helical repeat and linking number of surface-wrapped DNA. Science 241:323–327
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Liu, Y., Hua, ZC., Leng, F. (2018). DNA Supercoiling Measurement in Bacteria. In: Drolet, M. (eds) DNA Topoisomerases. Methods in Molecular Biology, vol 1703. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7459-7_4
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DOI: https://doi.org/10.1007/978-1-4939-7459-7_4
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Publisher Name: Humana Press, New York, NY
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Online ISBN: 978-1-4939-7459-7
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