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Microbiology 144 (1998), 3097-3104; DOI  10.1099/00221287-144-11-3097
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A vector for systematic gene inactivation in Bacillus subtilis

Valerie Vagner1,2, Etienne Dervyn1 and S. Dusko Ehrlich1

Genetique Microbienne, lnstitut National de la Recherche Ag ronom ique, Domaine de Vilvefl, 78352 Jouy-en-Josas cedex, France

2Author for correspondence: Valirie Vagner. Tel: +33 1 34 65 25 34. Fax: +33 1 34 65 25 21.e-mail: vagner@ biotec.jouy.inra.fr

ABSTRACT

SUMMARY: To study the functions of the uncharacterized open reading frames identified in the Bacillus subtih genome, several vectors were constructed t o perform insertional mutagenesis in the chromosome. All the pMUTlN plasmids carry a lac2 reporter gene and an inducible Pspac promoter, which is tightly regulated and tan be induced about 1000-fold. The integration of a pMUTlN vector into the target gene has three consequences: (1) the target gene is inactivated; (2) lac2 becomes transcriptionally fused t o the gene, allowing its expression pattern to be monitored; (3) the Pspac promoter controls the transcription of downstream genes in an IPTG-dependent fashion. This last feature is important because B. subti/is genes are often organized in operons. The potential polar effects generated by the integration of the vectors can be alleviated by addition of IPTG. Also, conditional mutants of essential genes can be obtained by integrating pMUTlN vectors upstream of the target gene. The vectors are currently being used for systematic inactivation of genes without known function within the B. subtilis European consortium. pMUTlN characteristics and the inactivation of eight genes in the resA-serA region of the chromosome are presented.


Keywords: integrational vector, Bacillus subtilis, inducible promoter, essential genes




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The H2O2 Stress-Responsive Regulator PerR Positively Regulates srfA Expression in Bacillus subtilis
J. Bacteriol., October 1, 2005; 187(19): 6659 - 6667.
[Abstract] [Full Text] [PDF]


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Proc. Natl. Acad. Sci. USAHome page
I. Gusarov and E. Nudler
NO-mediated cytoprotection: Instant adaptation to oxidative stress in bacteria
PNAS, September 27, 2005; 102(39): 13855 - 13860.
[Abstract] [Full Text] [PDF]


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J. Bacteriol.Home page
R. Ohki, K. Tateno, T. Takizawa, T. Aiso, and M. Murata
Transcriptional Termination Control of a Novel ABC Transporter Gene Involved in Antibiotic Resistance in Bacillus subtilis
J. Bacteriol., September 1, 2005; 187(17): 5946 - 5954.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
R. Bernard, M. El Ghachi, D. Mengin-Lecreulx, M. Chippaux, and F. Denizot
BcrC from Bacillus subtilis Acts as an Undecaprenyl Pyrophosphate Phosphatase in Bacitracin Resistance
J. Biol. Chem., August 12, 2005; 280(32): 28852 - 28857.
[Abstract] [Full Text] [PDF]


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J. Bacteriol.Home page
H. Szurmant, K. Nelson, E.-J. Kim, M. Perego, and J. A. Hoch
YycH Regulates the Activity of the Essential YycFG Two-Component System in Bacillus subtilis
J. Bacteriol., August 1, 2005; 187(15): 5419 - 5426.
[Abstract] [Full Text] [PDF]


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J. Bacteriol.Home page
K. Bailey-Smith, S. J. Todd, T. W. Southworth, J. Proctor, and A. Moir
The ExsA Protein of Bacillus cereus Is Required for Assembly of Coat and Exosporium onto the Spore Surface
J. Bacteriol., June 1, 2005; 187(11): 3800 - 3806.
[Abstract] [Full Text] [PDF]


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MicrobiologyHome page
M. Serizawa and J. Sekiguchi
The Bacillus subtilis YdfHI two-component system regulates the transcription of ydfJ, a member of the RND superfamily
Microbiology, June 1, 2005; 151(6): 1769 - 1778.
[Abstract] [Full Text] [PDF]


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MicrobiologyHome page
M. Pietiainen, M. Gardemeister, M. Mecklin, S. Leskela, M. Sarvas, and V. P. Kontinen
Cationic antimicrobial peptides elicit a complex stress response in Bacillus subtilis that involves ECF-type sigma factors and two-component signal transduction systems
Microbiology, May 1, 2005; 151(5): 1577 - 1592.
[Abstract] [Full Text] [PDF]


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J. Bacteriol.Home page
D. Hoper, U. Volker, and M. Hecker
Comprehensive Characterization of the Contribution of Individual SigB-Dependent General Stress Genes to Stress Resistance of Bacillus subtilis
J. Bacteriol., April 15, 2005; 187(8): 2810 - 2826.
[Abstract] [Full Text] [PDF]


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Appl. Environ. Microbiol.Home page
H. Westers, P. G. Braun, L. Westers, H. Antelmann, M. Hecker, J. D. H. Jongbloed, H. Yoshikawa, T. Tanaka, J. M. van Dijl, and W. J. Quax
Genes Involved in SkfA Killing Factor Production Protect a Bacillus subtilis Lipase against Proteolysis
Appl. Envir. Microbiol., April 1, 2005; 71(4): 1899 - 1908.
[Abstract] [Full Text] [PDF]


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J. Bacteriol.Home page
M. Albano, W. K. Smits, L. T. Y. Ho, B. Kraigher, I. Mandic-Mulec, O. P. Kuipers, and D. Dubnau
The Rok Protein of Bacillus subtilis Represses Genes for Cell Surface and Extracellular Functions
J. Bacteriol., March 15, 2005; 187(6): 2010 - 2019.
[Abstract] [Full Text] [PDF]


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MicrobiologyHome page
D.-J. Scheffers
Dynamic localization of penicillin-binding proteins during spore development in Bacillus subtilis
Microbiology, March 1, 2005; 151(3): 999 - 1012.
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Appl. Environ. Microbiol.Home page
L. M. Hornstra, Y. P. de Vries, W. M. de Vos, T. Abee, and M. H. J. Wells-Bennik
gerR, a Novel ger Operon Involved in L-Alanine- and Inosine-Initiated Germination of Bacillus cereus ATCC 14579
Appl. Envir. Microbiol., February 1, 2005; 71(2): 774 - 781.
[Abstract] [Full Text] [PDF]


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J. Bacteriol.Home page
P. Nygaard and H. H. Saxild
The Purine Efflux Pump PbuE in Bacillus subtilis Modulates Expression of the PurR and G-Box (XptR) Regulons by Adjusting the Purine Base Pool Size
J. Bacteriol., January 15, 2005; 187(2): 791 - 794.
[Abstract] [Full Text] [PDF]


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J. Bacteriol.Home page
D. Julkowska, M. Obuchowski, I. B. Holland, and Simone. J. Seror
Comparative Analysis of the Development of Swarming Communities of Bacillus subtilis 168 and a Natural Wild Type: Critical Effects of Surfactin and the Composition of the Medium
J. Bacteriol., January 1, 2005; 187(1): 65 - 76.
[Abstract] [Full Text] [PDF]


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J. Bacteriol.Home page
C. Sao-Jose, C. Baptista, and M. A. Santos
Bacillus subtilis Operon Encoding a Membrane Receptor for Bacteriophage SPP1
J. Bacteriol., December 15, 2004; 186(24): 8337 - 8346.
[Abstract] [Full Text] [PDF]


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J. Bacteriol.Home page
S. E. Dale, M. T. Sebulsky, and D. E. Heinrichs
Involvement of SirABC in Iron-Siderophore Import in Staphylococcus aureus
J. Bacteriol., December 15, 2004; 186(24): 8356 - 8362.
[Abstract] [Full Text] [PDF]


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Appl. Environ. Microbiol.Home page
M. Arnaud, A. Chastanet, and M. Debarbouille
New Vector for Efficient Allelic Replacement in Naturally Nontransformable, Low-GC-Content, Gram-Positive Bacteria
Appl. Envir. Microbiol., November 1, 2004; 70(11): 6887 - 6891.
[Abstract] [Full Text] [PDF]


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J. Bacteriol.Home page
G. Amati, P. Bisicchia, and A. Galizzi
DegU-P Represses Expression of the Motility fla-che Operon in Bacillus subtilis
J. Bacteriol., September 15, 2004; 186(18): 6003 - 6014.
[Abstract] [Full Text] [PDF]


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J. Bacteriol.Home page
K.-i. Yoshida, Y.-h. Ohki, M. Murata, M. Kinehara, H. Matsuoka, T. Satomura, R. Ohki, M. Kumano, K. Yamane, and Y. Fujita
Bacillus subtilis LmrA Is a Repressor of the lmrAB and yxaGH Operons: Identification of Its Binding Site and Functional Analysis of lmrB and yxaGH
J. Bacteriol., September 1, 2004; 186(17): 5640 - 5648.
[Abstract] [Full Text] [PDF]


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MicrobiologyHome page
K. Kimura, L.-S. P. Tran, and Y. Itoh
Roles and regulation of the glutamate racemase isogenes, racE and yrpC, in Bacillus subtilis
Microbiology, September 1, 2004; 150(9): 2911 - 2920.
[Abstract] [Full Text] [PDF]


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Antimicrob. Agents Chemother.Home page
T. Mascher, S. L. Zimmer, T.-A. Smith, and J. D. Helmann
Antibiotic-Inducible Promoter Regulated by the Cell Envelope Stress-Sensing Two-Component System LiaRS of Bacillus subtilis
Antimicrob. Agents Chemother., August 1, 2004; 48(8): 2888 - 2896.
[Abstract] [Full Text] [PDF]


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MicrobiologyHome page
N. E. E. Allenby, N. O'Connor, Z. Pragai, N. M. Carter, M. Miethke, S. Engelmann, M. Hecker, A. Wipat, A. C. Ward, and C. R. Harwood
Post-transcriptional regulation of the Bacillus subtilis pst operon encoding a phosphate-specific ABC transporter
Microbiology, August 1, 2004; 150(8): 2619 - 2628.
[Abstract] [Full Text] [PDF]


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J. Bacteriol.Home page
N. Zamboni, E. Fischer, D. Laudert, S. Aymerich, H.-P. Hohmann, and U. Sauer
The Bacillus subtilis yqjI Gene Encodes the NADP+-Dependent 6-P-Gluconate Dehydrogenase in the Pentose Phosphate Pathway
J. Bacteriol., July 15, 2004; 186(14): 4528 - 4534.
[Abstract] [Full Text] [PDF]


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Appl. Environ. Microbiol.Home page
M. Ashiuchi, K. Shimanouchi, H. Nakamura, T. Kamei, K. Soda, C. Park, M.-H. Sung, and H. Misono
Enzymatic Synthesis of High-Molecular-Mass Poly-{gamma}-Glutamate and Regulation of Its Stereochemistry
Appl. Envir. Microbiol., July 1, 2004; 70(7): 4249 - 4255.
[Abstract] [Full Text] [PDF]


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J. Bacteriol.Home page
S. S. Branda, J. E. Gonzalez-Pastor, E. Dervyn, S. D. Ehrlich, R. Losick, and R. Kolter
Genes Involved in Formation of Structured Multicellular Communities by Bacillus subtilis
J. Bacteriol., June 15, 2004; 186(12): 3970 - 3979.
[Abstract] [Full Text] [PDF]


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J. Bacteriol.Home page
B. Gorke, L. Fraysse, and A. Galinier
Drastic Differences in Crh and HPr Synthesis Levels Reflect Their Different Impacts on Catabolite Repression in Bacillus subtilis
J. Bacteriol., May 15, 2004; 186(10): 2992 - 2995.
[Abstract] [Full Text] [PDF]


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J. Bacteriol.Home page
M. Ogura, A. Matsuzawa, H. Yoshikawa, and T. Tanaka
Bacillus subtilis SalA (YbaL) Negatively Regulates Expression of scoC, Which Encodes the Repressor for the Alkaline Exoprotease Gene, aprE
J. Bacteriol., May 15, 2004; 186(10): 3056 - 3064.
[Abstract] [Full Text] [PDF]


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MicrobiologyHome page
K.-i. Yoshida, M. Yamaguchi, H. Ikeda, K. Omae, K.-i. Tsurusaki, and Y. Fujita
The fifth gene of the iol operon of Bacillus subtilis, iolE, encodes 2-keto-myo-inositol dehydratase
Microbiology, March 1, 2004; 150(3): 571 - 580.
[Abstract] [Full Text] [PDF]


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MicrobiologyHome page
M. Yoshimura, K. Asai, Y. Sadaie, and H. Yoshikawa
Interaction of Bacillus subtilis extracytoplasmic function (ECF) sigma factors with the N-terminal regions of their potential anti-sigma factors
Microbiology, March 1, 2004; 150(3): 591 - 599.
[Abstract] [Full Text] [PDF]


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J. Bacteriol.Home page
M. Cao and J. D. Helmann
The Bacillus subtilis Extracytoplasmic-Function {sigma}X Factor Regulates Modification of the Cell Envelope and Resistance to Cationic Antimicrobial Peptides
J. Bacteriol., February 15, 2004; 186(4): 1136 - 1146.
[Abstract] [Full Text] [PDF]




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