Expression, purification, and DNA-binding activity of the Herbaspirillum seropedicae RecX protein

Carolina W Galvão; Fábio O Pedrosa; Emanuel M Souza; M Geoffrey Yates; Leda S Chubatsu; Maria Berenice R Steffens

doi:10.1016/j.pep.2004.01.014

Expression, purification, and DNA-binding activity of the Herbaspirillum seropedicae RecX protein

Protein Expr Purif. 2004 Jun;35(2):298-303. doi: 10.1016/j.pep.2004.01.014.

Authors

Carolina W Galvão¹, Fábio O Pedrosa, Emanuel M Souza, M Geoffrey Yates, Leda S Chubatsu, Maria Berenice R Steffens

Affiliation

¹ Department of Biochemistry and Molecular Biology, Universidade Federal do Paraná, C.P. 19046, Curitiba, PR 81531-990, Brazil.

PMID: 15135406
DOI: 10.1016/j.pep.2004.01.014

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Bacterial Proteins / genetics
Bacterial Proteins / isolation & purification
Bacterial Proteins / metabolism*
Base Sequence
Chromatography, Affinity
DNA Primers
Herbaspirillum / metabolism*

Substances

Bacterial Proteins
DNA Primers
RecX protein, Xanthomonas campestris