Abstract
To perform mechanistic studies on the biology of bacteria including metabolism, physiology, and pathogenesis, it is essential to possess the tools required for genetic manipulation. Introduction of plasmid DNA into Staphylococcus epidermidis for subsequent genetic manipulation, including allelic replacement and complementation experiments, is typically performed by either electroporation or conjugative mobilization. Herein, standard protocols and tips for the transfer of plasmid DNA to S. epidermidis by these two methods are provided.
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References
Griffith F (1928) The significance of pneumococcal types. J Hyg (Lond) 27:113–159
Lindberg M, Sjostrom J-E, Johansson T (1972) Transformation of chromosomal and plasmid characters in Staphylococcus aureus. J Bacteriol 109:844–847
Nomura H, Udou T, Yoshida K et al (1971) Induction of hemolysin synthesis by transformation in Staphylococcus aureus. J Bacteriol 105:673–675
Thompson NE, Pattee PA (1977) Transformation in Staphylococcus aureus: role of bacteriophage and incidence of competence among strains. J Bacteriol 129:778–788
Thompson NE, Pattee PA (1981) Genetic transformation in Staphylococcus aureus: demonstration of a competence-conferring factor of bacteriophage origin in bacteriophage 80α lysates. J Bacteriol 148:294–300
Birmingham VA, Pattee PA (1981) Genetic transformation in Staphylococcus aureus: isolation and characterization of a competence-conferring factor from bacteriophage 80α lysates. J Bacteriol 148:301–307
Morikawa K, Takemura AJ, Inose Y et al (2012) Expression of a cryptic secondary sigma factor gene unveils natural competence for DNA transformation in Staphylococcus aureus. PLoS Pathog 8:1–20
Gotz F, Schumacher B (1987) Improvements of protoplast transformation in Staphylococcus carnosus. FEMS Microbiol Lett 40:285–288
Augustin J, Gotz F (1990) Transformation of Staphylococcus epidermidis and other staphylococcal species with plasmid DNA by electroporation. FEMS Microbiol Lett 66:203–207
Waldron DE, Lindsay JA (2006) Sau1: a novel lineage-specific type I restriction-modification system that blocks horizontal gene transfer into Staphylococcus aureus and between S. aureus isolates of different lineages. J Bacteriol 188:5578–5585
Corvaglia AR, Francois P, Hernandez D et al (2010) A type III-like restriction endonuclease functions as a major barrier to horizontal gene transfer in clinical Staphylococcus aureus strains. Proc Natl Acad Sci U S A 107:11954–11958
Xu SY, Corvaglia AR, Chan SH et al (2011) A type IV modification-dependent restriction enzyme SauUSI from Staphylococcus aureus subsp. aureus USA300. Nucleic Acids Res 39:5597–5610
Monk IR, Shah IM, Xu M et al (2012) Transforming the untransformable: application of direct transformation to manipulate genetically Staphylococcus aureus and Staphylococcus epidermidis. MBio 3:1–11
Marraffini LA, Sontheimer EJ (2008) CRISPR interference limits horizontal gene transfer in staphylococci by targeting DNA. Science 322:1843–1845
Archer GL, Johnston JL (1983) Self-transmissable plasmids in staphylococci that encode resistance to aminoglycosides. Antimicrob Agents Chemother 24:70–77
Forbes BA, Schaberg DR (1983) Transfer of resistance plasmids from Staphylococcus epidermidis to Staphylococcus aureus: evidence for conjugative exchange of resistance. J Bacteriol 153:627–634
Jaffe HW, Sweeney HM, Nathan C et al (1980) Identity and interspecific transfer of gentamicin-resistance plasmids in Staphylococcus aureus and Staphylococcus epidermidis. J Infect Dis 141:738–747
Caryl JA, O’Neill AJ (2009) Complete nucleotide sequence of pGO1, the prototype conjugative plasmid from the Staphylococci. Plasmid 62:35–38
Thomas WD, Archer G (1989) Identification and cloning of the conjugative transfer region of Staphylococcus aureus plasmid pGO1. J Bacteriol 171:684–691
Projan SJ, Archer GL (1989) Mobilization of the relaxable Staphylococcus aureus plasmid pC221 by the conjugative plasmid pGO1 involves three pC22 loci. J Bacteriol 171:1941–1845
Novick R (1976) Plasmid-protein relaxation complexes in Staphylococcus aureus. J Bacteriol 127:1177–1187
Caryl JA, Smith MCA, Thomas CD (2004) Reconstitution of a staphylococcal plasmid-protein relaxation complex in vitro. J Bacteriol 186:3374–3383
Mack D, Fischer W, Krokotsch A et al (1996) The intercellular adhesin involved in biofilm accumulation of Staphylococcus epidermidis is a linear beta-1,6-linked glucosaminoglycan: purification and structural analysis. J Bacteriol 178:175–183
Aubry-Damon H, Soussy C-J, Courvalin P (1998) Characterization of mutations in the rpoB gene that confer rifampin resistance in Staphylococcus aureus. Antimicrob Agents Chemother 42:2590–2594
Stieger M, Angehrn P, Wohlgensinger B et al (1996) GyrB mutations in Staphylococcus aureus strains resistant to cyclothialidine, coumermycin, and novobiocin. Antimicrob Agents Chemother 40:1060–1062
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Maliszewski, K.L., Nuxoll, A.S. (2014). Use of Electroporation and Conjugative Mobilization for Genetic Manipulation of Staphylococcus epidermidis . In: Fey, P. (eds) Staphylococcus Epidermidis. Methods in Molecular Biology, vol 1106. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-736-5_11
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DOI: https://doi.org/10.1007/978-1-62703-736-5_11
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-735-8
Online ISBN: 978-1-62703-736-5
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