Factors and Conditions That Impact Electroporation of Clostridioides difficile Strains.

MSphere
Disha Bhattacharjee, Joseph A Sorg

Abstract

An important risk factor for acquiring Clostridioides difficile infection is antibiotic use. Therefore, a detailed knowledge of the physiology and the virulence factors can help drive the development of new diagnostic tools and nonantibiotic therapeutic agents to combat these organisms. Several genetic systems are available to study C. difficile in the laboratory environment, and all rely on stably replicating or segregationally unstable plasmids. Currently, the transfer of plasmids into C. difficile can only be performed by conjugation using Escherichia coli or Bacillus subtilis as conjugal donors. Here we report a method to introduce plasmid DNA into C. difficile using electroporation and test factors that might contribute to higher transformation efficiencies: osmolyte used to stabilize weakened cells, DNA concentration, and recovery time postelectroporation. Depending on the C. difficile strain and plasmid used, this transformation protocol achieves between 20 and 200 colonies per microgram of DNA and is mostly influenced by the recovery time postelectroporation. Based on our findings, we recommend that each strain be tested for the optimum recovery time in each lab.IMPORTANCE Understanding the underlying biology of pathoge...Continue Reading

References

Apr 1, 1991·Applied and Environmental Microbiology·G M DunnyD J LeBlanc
Jan 1, 1991·Methods in Enzymology·D M Becker, L Guarente
Feb 1, 1988·Proceedings of the National Academy of Sciences of the United States of America·J F MillerL S Tompkins
Jun 1, 1988·Journal of Bacteriology·N M Calvin, P C Hanawalt
Nov 1, 1973·Journal of Bacteriology·W HammesO Kandler
Jan 1, 1981·Molecular & General Genetics : MGG·W M de VosT A Trautner
Aug 15, 1997·FEMS Microbiology Letters·F ArtiguenaveC Danglot
Dec 29, 1998·Applied Microbiology and Biotechnology·S NakotteP Dürre
Jun 1, 2000·Proceedings of the National Academy of Sciences of the United States of America·K A Datsenko, B L Wanner
Dec 2, 2000·Microbiology·K C JennertM Young
Dec 9, 2003·FEMS Microbiology Letters·Michael HerbertNigel P Minton
Apr 13, 2005·Journal of Microbiological Methods·Alena JiráskováPavel Branny
Sep 21, 2005·Applied Microbiology and Biotechnology·L BroettoI S Schrank
Jun 1, 1992·Plant Physiology·S L Van Wert, J A Saunders
Jul 31, 2007·Journal of Microbiological Methods·John T HeapNigel P Minton
Feb 12, 2008·Current Protocols in Molecular Biology·Huntington Potter
Dec 1, 2009·Applied Microbiology and Biotechnology·Trond Erik Vee Aune, Finn Lillelund Aachmann
Jul 10, 2010·Proceedings of the National Academy of Sciences of the United States of America·Michael KöpkePeter Dürre
Jun 11, 2011·The Journal of Biological Chemistry·Robert P Fagan, Neil F Fairweather
May 23, 2012·Methods in Enzymology·Daniel G Olson, Lee R Lynd
Jul 4, 2012·Bioengineered·Sarah A Kuehne, Nigel P Minton
Apr 11, 2013·Biotechnology for Biofuels·Michael E PyneC Perry Chou
Feb 8, 2014·Gastroenterology·Robert A Britton, Vincent B Young
Feb 26, 2015·The New England Journal of Medicine·Fernanda C LessaL Clifford McDonald
Apr 4, 2015·International Journal of Molecular Sciences·Zhi ZhangHong Tan
Nov 5, 2016·Anaerobe·Joseph A Kirk, Robert P Fagan
Jan 4, 2017·Journal of Microbiological Methods·Yanglei Yi, Oscar P Kuipers
Jan 13, 2017·International Journal of Systematic and Evolutionary Microbiology·Aharon Oren, George M Garrity
Nov 6, 2018·Frontiers in Microbiology·Disha Bhattacharjee, Joseph A Sorg
Aug 11, 2019·Applied and Environmental Microbiology·Anna MaikovaOlga Soutourina

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Citations

Apr 7, 2021·Journal of Bacteriology·Kathleen N McAllisterJoseph A Sorg

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