Mobilizable Plasmids for Tunable Gene Expression in Francisella novicida

Frontiers in Cellular and Infection Microbiology
Maj BrodmannMarek Basler

Abstract

Francisella tularensis is the causative agent of the life-threatening disease tularemia. However, the molecular tools to study Francisella are limited. Especially, expression plasmids are sparse and difficult to use, as they are unstable and prone to spontaneous loss. Most Francisella expression plasmids lack inducible promoters making it difficult to control gene expression levels. In addition, available expression plasmids are mainly designed for F. tularensis, however, genetic differences including restriction-modification systems impede the use of these plasmids in F. novicida, which is often used as a model organism to study Francisella pathogenesis. Here we report construction and characterization of two mobilizable plasmids (pFNMB1 and pFNMB2) designed for regulated gene expression in F. novicida. pFNMB plasmids contain a tetracycline inducible promoter to control gene expression levels and oriT for RP4 mediated mobilization. We show that both plasmids are stably maintained in bacteria for more than 40 generations over 4 days of culturing in the absence of selection against plasmid loss. Expression levels are dependent on anhydrotetracycline concentration and homogeneous in a bacterial population. pFNMB1 and pFNMB2 plasm...Continue Reading

References

Nov 1, 1976·Proceedings of the National Academy of Sciences of the United States of America·B PoliskyD H Gelfand
Jun 15, 1992·Proceedings of the National Academy of Sciences of the United States of America·M Gossen, H Bujard
Dec 1, 1991·Journal of General Microbiology·L S AnthonyF E Nano
Mar 1, 1996·FEMS Immunology and Medical Microbiology·A NorqvistG Sandström
Dec 12, 2001·Plasmid·A P PomerantsevV M Pavlov
Jan 7, 2003·Applied and Environmental Microbiology·Hadi AbdMats Forsman
May 29, 2003·FEMS Microbiology Letters·Igor GolovliovVitaly Pavlov
Jul 17, 2004·Molecular Microbiology·Oliver ScholzWolfgang Hillen
Nov 20, 2004·Nature Reviews. Microbiology·Petra C F OystonRichard W Titball
Dec 3, 2004·Applied and Environmental Microbiology·Tamara M MaierThomas C Zahrt
Jan 11, 2005·Nature Genetics·Pär LarssonRichard W Titball
Mar 7, 2006·Applied and Environmental Microbiology·Tamara M MaierD W Frank
Aug 16, 2006·Proceedings of the National Academy of Sciences of the United States of America·Horacio GilDavid G Thanassi
Aug 24, 2006·Infection and Immunity·Rebecca TempelFred Heffron
Mar 29, 2007·Proceedings of the National Academy of Sciences of the United States of America·David S WeissDenise M Monack
Mar 31, 2007·Annals of the New York Academy of Sciences·Francis E Nano, Crystal Schmerk
Mar 31, 2007·Annals of the New York Academy of Sciences·Dara W Frank, Thomas C Zahrt
Feb 5, 2008·Applied and Environmental Microbiology·Joseph HorzempaGerard J Nau
Oct 7, 2008·Journal of Bacteriology·Larry A GallagherColin Manoil
Dec 11, 2008·FEMS Microbiology Letters·Eric D LoVulloMartin S Pavelka
Mar 31, 2010·Nature Immunology·Teresa Fernandes-AlnemriEmad S Alnemri
May 12, 2010·Proceedings of the National Academy of Sciences of the United States of America·Jonathan W JonesDenise M Monack
Jan 1, 2008·Microbial Biotechnology·Ralph Bertram, Wolfgang Hillen
May 25, 2011·Microbial Pathogenesis·Nicole M Ark, Barbara J Mann
Jan 1, 2010·Frontiers in Microbiology·Xhavit Zogaj, Karl E Klose
Jul 13, 2011·Frontiers in Microbiology·Marina SanticYousef Abu Kwaik
Jun 30, 2012·Nature Methods·Johannes SchindelinAlbert Cardona
Jul 24, 2012·Applied and Environmental Microbiology·Eric D LoVulloThomas H Kawula
Aug 22, 2013·The Journal of Antimicrobial Chemotherapy·Vivien SuteraMax Maurin
Mar 25, 2014·Frontiers in Cellular and Infection Microbiology·Luke C Kingry, Jeannine M Petersen
Feb 28, 2015·Cell·Mikhail KudryashevMarek Basler

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Citations

Jun 1, 2021·Frontiers in Microbiology·Stephen J Kassinger, Monique L van Hoek
Oct 30, 2021·Microbial Biotechnology·Ralph BertramChristopher F Schuster

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Methods Mentioned

BETA
RepA
PCR
FCS
fluorescence microscopy

Software Mentioned

Fiji

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