In vivo Himar1-based transposon mutagenesis of Francisella tularensis

Applied and Environmental Microbiology
Tamara M MaierDara W Frank

Abstract

Francisella tularensis is the intracellular pathogen that causes human tularemia. It is recognized as a potential agent of bioterrorism due to its low infectious dose and multiple routes of entry. We report the development of a Himar1-based random mutagenesis system for F. tularensis (HimarFT). In vivo mutagenesis of F. tularensis live vaccine strain (LVS) with HimarFT occurs at high efficiency. Approximately 12 to 15% of cells transformed with the delivery plasmid result in transposon insertion into the genome. Results from Southern blot analysis of 33 random isolates suggest that single insertions occurred, accompanied by the loss of the plasmid vehicle in most cases. Nucleotide sequence analysis of rescued genomic DNA with HimarFT indicates that the orientation of integration was unbiased and that insertions occurred in open reading frames and intergenic and repetitive regions of the chromosome. To determine the utility of the system, transposon mutagenesis was performed, followed by a screen for growth on Chamberlain's chemically defined medium (CDM) to isolate auxotrophic mutants. Several mutants were isolated that grew on complex but not on the CDM. We genetically complemented two of the mutants for growth on CDM with a n...Continue Reading

References

Dec 1, 1991·Journal of General Microbiology·L S AnthonyF E Nano
Sep 1, 1991·Infection and Immunity·A H FortierC A Nacy
Sep 1, 1991·Infection and Immunity·L D AnthonyF E Nano
Aug 1, 1986·Journal of Bacteriology·M A Vandeyar, S A Zahler
Oct 1, 1994·The Journal of Infectious Diseases·N BhatnagarA Fortier
May 3, 1996·The Journal of Biological Chemistry·T J ReillyM S Kuhlenschmidt
Oct 1, 1996·The EMBO Journal·D J LampeH M Robertson
Jan 1, 1997·Annual Review of Biochemistry·N L Craig
Nov 14, 1997·Letters in Applied Microbiology·P Engel, A R Lax
Jul 22, 1998·Proceedings of the National Academy of Sciences of the United States of America·B J AkerleyJ J Mekalanos
Aug 14, 1998·Molecular Microbiology·G S Baron, F E Nano
Feb 17, 1999·Proceedings of the National Academy of Sciences of the United States of America·E J RubinJ J Mekalanos
Sep 15, 2000·Journal of Bacteriology·V PelicicX Nassif
Dec 12, 2001·Plasmid·A P PomerantsevV M Pavlov
Jan 24, 2002·Proceedings of the National Academy of Sciences of the United States of America·Brian J AkerleyJohn J Mekalanos
Oct 24, 2002·FEMS Microbiology Letters·Catherine G GrayFrancis E Nano
Jan 7, 2003·Applied and Environmental Microbiology·Hadi AbdMats Forsman
May 29, 2003·FEMS Microbiology Letters·Igor GolovliovVitaly Pavlov
Jul 23, 2003·Journal of Basic Microbiology·Tamara M MaierCharles R Myers
Nov 18, 2003·Proceedings of the National Academy of Sciences of the United States of America·Michael A JacobsColin Manoil
Dec 19, 2003·FEMS Microbiology Letters·Crystal M LaurianoKarl E Klose
Sep 11, 2004·Journal of Medical Microbiology·Helena LindgrenAnders Sjöstedt
Sep 18, 2004·Journal of Bacteriology·Francis E NanoKaren L Elkins
Sep 24, 2004·Infection and Immunity·Jochen StritzkerWerner Goebel
Oct 7, 2004·Applied and Environmental Microbiology·Philip E StewartPatricia A Rosa
Nov 6, 2004·Applied and Environmental Microbiology·Thomas H KawulaRobin R Craven
Dec 3, 2004·Applied and Environmental Microbiology·Tamara M MaierThomas C Zahrt
Jan 11, 2005·Nature Genetics·Pär LarssonRichard W Titball
May 28, 2005·Advanced Drug Delivery Reviews·Karen E IsherwoodW John W Morrow

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Citations

Nov 13, 2012·Applied Microbiology and Biotechnology·Bohdan BilykAndriy Luzhetskyy
Aug 30, 2008·Applied and Environmental Microbiology·Yu YangChunhao Li
Oct 28, 2008·Applied and Environmental Microbiology·Drew A RhollHerbert P Schweizer
Dec 2, 2009·Applied and Environmental Microbiology·Jeffrey J MargolisDenise M Monack
Feb 5, 2008·Applied and Environmental Microbiology·Joseph HorzempaGerard J Nau
Dec 22, 2009·Applied and Environmental Microbiology·Stephen T Cartman, Nigel P Minton
Mar 27, 2013·Infection and Immunity·Lydia M BarriganJeffrey A Frelinger
Apr 11, 2007·Infection and Immunity·Jingliang SuJing-Ren Zhang
Dec 1, 2009·Microbiology and Molecular Biology Reviews : MMBR·Roger D PechousThomas C Zahrt
Nov 2, 2006·BMC Biotechnology·Roderick F FelsheimUlrike G Munderloh
Aug 16, 2006·Proceedings of the National Academy of Sciences of the United States of America·Horacio GilDavid G Thanassi
Oct 22, 2013·Applied and Environmental Microbiology·Ralph L McWhinnie, Francis E Nano
May 25, 2011·Microbial Pathogenesis·Nicole M Ark, Barbara J Mann
Sep 12, 2015·IEEE/ACM Transactions on Computational Biology and Bioinformatics·Michael A DeJesus, Thomas R Ioerger
Jul 21, 2015·Molecular Microbiology·Terry BrissacAlain Charbit
Nov 30, 2013·Cellular Microbiology·Elizabeth Di Russo CaseJean Celli
Jul 17, 2008·Journal of Medical Microbiology·Petra C F Oyston
Apr 1, 2009·Microbiology·Eric D LoVulloMartin S Pavelka
May 16, 2009·Microbiology·Nathalie GrallKarin L Meibom
Jan 12, 2007·Proceedings of the National Academy of Sciences of the United States of America·Larry A GallagherColin Manoil
Feb 24, 2007·FEMS Immunology and Medical Microbiology·Kate F GriffinRichard W Titball
Dec 11, 2008·FEMS Microbiology Letters·Eric D LoVulloMartin S Pavelka
Nov 21, 2007·FEMS Microbiology Letters·Jagjit S LuduFrancis E Nano
Jul 28, 2016·Journal of Microbiological Methods·Stephanie S LehmanHerbert P Schweizer
May 15, 2013·Molecular & Cellular Proteomics : MCP·Jennifer DieppedaleAlain Charbit

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