Expression, purification, and DNA-binding activity of the Herbaspirillum seropedicae RecX protein.
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.
Galvão
Carolina W
CW
Department of Biochemistry and Molecular Biology, Universidade Federal do Paraná, C.P. 19046, Curitiba, PR 81531-990, Brazil.
Pedrosa
Fábio O
FO
Souza
Emanuel M
EM
Yates
M Geoffrey
MG
Chubatsu
Leda S
LS
Steffens
Maria Berenice R
MB
D001426:Bacterial Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D001483:Base Sequence; D002846:Chromatography, Affinity; D017931:DNA Primers; D042803:Herbaspirillum / Q000378:metabolism*
D001426:Bacterial Proteins; D017931:DNA Primers; C430523:RecX protein, Xanthomonas campestris