PMID: 8606153Apr 1, 1996Paper

In vitro binding of the Salmonella dublin virulence plasmid regulatory protein SpvR to the promoter regions of spvA and spvR

Journal of Bacteriology
P Grob, D G Guiney

Abstract

The spv regulon of Salmonella dublin is essential for virulence in mice. SpvR, a LysR-type regulator, induces the expression of the spvABCD operon and its own expression in the stationary phase of bacterial growth and in macrophages. We constructed fusion proteins to the maltose-binding protein (MBP) and a His tag peptide (His) to overcome the insolubility and to facilitate purification of SpvR. We demonstrated that both fusion proteins, MBP-SpvR and His-SpvR, were able to induce spvA expression in vivo. MBP-SpvR was produced as soluble protein, whereas His-SpvR was only marginally present in the soluble cell fraction. Affinity chromatography resulted in at least 95% pure MBP-SpvR protein and in an enrichment of His-SpvR. Gel mobility shift assay revealed that the SpvR fusion proteins were able to bind to 125-and 147-bp DNA fragments of the spvA and spvR promoter regions, respectively. DNase I footprint experiments showed that the fusion proteins protected DNA regions of 54 and 50 bp within the spvA and spvR promoter regions, respectively.

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Citations

Dec 28, 2006·Nature Reviews. Microbiology·Charles J Dorman
Jan 16, 2014·Indian Journal of Microbiology·Abdollah DerakhshandehRahem Khoshbakht
Jun 10, 2004·Molecular Microbiology·Javier Pizarro-Cerdá, Karsten Tedin
Aug 26, 2018·Genetics·Ioannis PassarisAbram Aertsen
May 16, 2002·Infection and Immunity·Stephen J LibbyDonald G Guiney
Jan 20, 2010·Trends in Microbiology·Bethany N Kent, Seth R Bordenstein

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