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Expression, purification, and DNA-binding activity of the Herbaspirillum seropedicae RecX protein
Carolina W Galvão1, Fábio O Pedrosa, Emanuel M Souza
1Department of Biochemistry and Molecular Biology, Universidade Federal do Paraná, C.P. 19046, Curitiba, PR 81531-990, Brazil.
Abstract:
The Herbaspirillum seropedicae RecX protein participates in the SOS response: a process in which the RecA protein plays a central role. The RecX protein of the H. seropedicae, fused to a His-tag sequence (RecX His-tagged), was over-expressed in Escherichia coli and purified by metal-affinity chromatography to yield a highly purified and active protein. DNA band-shift assays showed that the RecX His-tagged protein bound to both circular and linear double-stranded DNA and also to circular single-stranded DNA. The apparent affinity of RecX for DNA decreased in the presence of Mg(2+) ions. The ability of RecX to bind DNA may be relevant to its function in the SOS response.
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