Gel-retardation assay of DNA-binding activity of the GyrA mutants in the presence of GyrB
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Taken from "The key DNA-binding residues in the C-terminal domain of DNA gyrase A subunit (GyrA)"
Nucleic Acids Research 2006;34(19):5650-5659.
Published online 11 Oct 2006
© 2006 The Author(s)All GyrA mutants were mixed with GyrB at an equal molar concentration (1.0 μM) at 25°C for ∼20 min, and incubated with 75 ng of 240 bp DNA in 20 μl DNA-binding buffer. Lane 1, DNA sample without any proteins; lane 2, GyrA alone; lanes 3–9, wild-type gyrase, E514A, Y577A, D669A, R691A, G729W, R745A, respectively plus DNA and GyrB; lane 10, GyrB only.