Vectors for regulated gene expression in the radioresistant bacterium Deinococcus radiodurans

Gene
François LecointeA Bailone

Abstract

Deinococcus radiodurans possesses an exceptional capacity to withstand the lethal and mutagenic effects of most form of DNA damage and has received considerable interest for use in both fundamental and applied research. Here we describe vectors that allow regulated expression of Deinococcal genes for functional analysis. The vectors contain the IPTG-regulated Spac system (Pspac promoter and lacI repressor gene), originally designed for Bacillus subtilis, that we have adapted to be functional in D. radiodurans. We show that the Spac system can control the expression of a lacZ reporter gene over two orders of magnitude depending on the inducer concentration and the copy number of the lacI regulatory gene. Furthermore, we demonstrate that the Spac system can be used to regulate the synthesis of a critical repair protein, such as RecA, resulting in a conditional mitomycin-resistant cell phenotype. We have also developed tools for the construction of conditional mutants where the expression of the target gene is regulated by an inducible promoter. The utility of these conditional gene inactivation systems is exemplified by the conditional lethal phenotype of a mutant expressing gyrA from the Pspac promoter.

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Citations

Oct 21, 2009·Proceedings of the National Academy of Sciences of the United States of America·Bhumit A PatelBrian R Crane
Oct 13, 2011·Molecular & Cellular Proteomics : MCP·Bhakti Basu, Shree Kumar Apte
Apr 29, 2008·Applied and Environmental Microbiology·Ian R MonkColin Hill
May 15, 2007·Journal of Bacteriology·Matthew D Servinsky, Douglas A Julin
Mar 5, 2011·Microbiology and Molecular Biology Reviews : MMBR·Dea Slade, Miroslav Radman
Apr 9, 2011·Standards in Genomic Sciences·Natalia IvanovaAlla Lapidus
Jun 9, 2009·Plasmid·Katsuya SatohIssay Narumi
Dec 20, 2005·Molecular Microbiology·Edmond JolivetSuzanne Sommer
Mar 7, 2013·Molecular Microbiology·Cécile PasternakSuzanne Sommer
May 20, 2015·Molecular Microbiology·Fanny Marie PassotSuzanne Sommer
Aug 7, 2018·Journal of Basic Microbiology·Harinder Singh
Jun 24, 2008·Critical Reviews in Biochemistry and Molecular Biology·Melanie BlasiusUlrich Hübscher
Sep 1, 2019·Applied and Environmental Microbiology·Angela ChenLydia M Contreras
May 27, 2021·Scientific Reports·Jelena ReparKsenija Zahradka

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