Abstract
A cornucopia of methods and molecular tools is available for genetic modification of staphylococci, as shown for at least ten different species to date (Prax et al. Microbiology 159:421–435, 2013). This chapter reviews a number of frequently used vectors for complementation purposes that usually replicate in E. coli and staphylococci and differ in parameters including copy number, mode of replication, and sequence length. Systems for the artificial control of gene expression are described that are modulated by low-molecular-weight effectors such as metal cations, carbohydrates, and antibiotics. Finally, the usefulness of reporter proteins that exhibit enzymatic or autofluorescent characteristics in staphylococci is highlighted.
Keywords:
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Prax M, Lee CY, Bertram R (2013) An update on the molecular genetics toolbox for staphylococci. Microbiology 159:421–435
McNamara P (2008) Genetic manipulation of Staphylococcus aureus. In: Lindsay J (ed) Staphylococcus molecular genetics. Caister Academic Press, Norfolk, UK
Novick RP (1989) Staphylococcal plasmids and their replication. Annu Rev Microbiol 43:537–565
Brückner R (1992) A series of shuttle vectors for Bacillus subtilis and Escherichia coli. Gene 122:187–192
Brückner R, Zyprian E, Matzura H (1984) Expression of a chloramphenicol-resistance determinant carried on hybrid plasmids in gram-positive and gram-negative bacteria. Gene 32:151–160
Firth N, Apisiridej S, Berg T et al (2000) Replication of staphylococcal multiresistance plasmids. J Bacteriol 182:2170–2178
Grkovic S, Brown MH, Hardie KM et al (2003) Stable low-copy-number Staphylococcus aureus shuttle vectors. Microbiology 149:785–794
Charpentier E, Anton AI, Barry P et al (2004) Novel cassette-based shuttle vector system for gram-positive bacteria. Appl Environ Microbiol 70:6076–6085
Corbisier P, Ji G, Nuyts G et al (1993) luxAB gene fusions with the arsenic and cadmium resistance operons of Staphylococcus aureus plasmid pI258. FEMS Microbiol Lett 110:231–238
Vandenesch F, Kornblum J, Novick RP (1991) A temporal signal, independent of agr, is required for hla but not spa transcription in Staphylococcus aureus. J Bacteriol 173:6313–6320
Hussain M, Becker K, von Eiff C et al (2001) Identification and characterization of a novel 38.5-kilodalton cell surface protein of Staphylococcus aureus with extended-spectrum binding activity for extracellular matrix and plasma proteins. J Bacteriol 183:6778–6786
Peschel A, Ottenwälder B, Götz F (1996) Inducible production and cellular location of the epidermin biosynthetic enzyme EpiB using an improved staphylococcal expression system. FEMS Microbiol Lett 137:279–284
Krismer B (1999) Studium der Funktion der sekretierten Proteine SceA und SceB, Analyse des Galaktoseoperons galRKET und Konstruktion von Sekretions- und Expressionsvektoren in Staphylococcus carnosus. PhD thesis, University of Tübingen, Tübingen
Wieland KP, Wieland B, Götz F (1995) A promoter-screening plasmid and xylose-inducible, glucose-repressible expression vectors for Staphylococcus carnosus. Gene 158:91–96
Sizemore C, Wieland B, Götz F et al (1992) Regulation of Staphylococcus xylosus xylose utilization genes at the molecular level. J Bacteriol 174:3042–3048
Hueck CJ, Hillen W, Saier MH Jr (1994) Analysis of a cis-active sequence mediating catabolite repression in gram-positive bacteria. Res Microbiol 145:503–518
Nega M, Dube L, Ziebandt AK et al (2014) Secretome analysis revealed adaptive and non-adaptive responses of the Staphylococcus carnosus femB mutant. Proteomics. doi: 10.1002/pmic.201400343
Yansura DG, Henner DJ (1984) Use of the Escherichia coli lac repressor and operator to control gene expression in Bacillus subtilis. Proc Natl Acad Sci U S A 81:439–443
Geissendörfer M, Hillen W (1990) Regulated expression of heterologous genes in Bacillus subtilis using the Tn10 encoded tet regulatory elements. Appl Microbiol Biotechnol 33:657–663
Zhang L, Fan F, Palmer LM et al (2000) Regulated gene expression in Staphylococcus aureus for identifying conditional lethal phenotypes and antibiotic mode of action. Gene 255:297–305
Bertram R, Hillen W (2008) The application of Tet repressor in prokaryotic gene regulation and expression. Microb Biotechnol 1:2–16
Ji Y, Marra A, Rosenberg M et al (1999) Regulated antisense RNA eliminates alpha-toxin virulence in Staphylococcus aureus infection. J Bacteriol 181:6585–6590
Gründling A, Schneewind O (2007) Genes required for glycolipid synthesis and lipoteichoic acid anchoring in Staphylococcus aureus. J Bacteriol 189:2521–2530
Gründling A, Schneewind O (2007) Synthesis of glycerol phosphate lipoteichoic acid in Staphylococcus aureus. Proc Natl Acad Sci U S A 104:8478–8483
Bateman BT, Donegan NP, Jarry TM et al (2001) Evaluation of a tetracycline-inducible promoter in Staphylococcus aureus in vitro and in vivo and its application in demonstrating the role of sigB in microcolony formation. Infect Immun 69:7851–7857
Corrigan RM, Foster TJ (2009) An improved tetracycline-inducible expression vector for Staphylococcus aureus. Plasmid 61:126–129
Helle L, Kull M, Mayer S et al (2011) Vectors for improved Tet repressor-dependent gradual gene induction or silencing in Staphylococcus aureus. Microbiology 157:3314–3323
Xu HH, Trawick JD, Haselbeck RJ et al (2010) Staphylococcus aureus TargetArray: comprehensive differential essential gene expression as a mechanistic tool to profile antibacterials. Antimicrob Agents Chemother 54:3659–3670
Stary E, Gaupp R, Lechner S et al (2010) New architectures for Tet-on and Tet-off regulation in Staphylococcus aureus. Appl Environ Microbiol 76:680–687
Kamionka A, Bogdanska-Urbaniak J, Scholz O et al (2004) Two mutations in the tetracycline repressor change the inducer anhydrotetracycline to a corepressor. Nucleic Acids Res 32:842–847
Scholz O, Henssler EM, Bail J et al (2004) Activity reversal of Tet repressor caused by single amino acid exchanges. Mol Microbiol 53:777–789
Schofield DA, Westwater C, Hoel BD et al (2003) Development of a thermally regulated broad-spectrum promoter system for use in pathogenic gram-positive species. Appl Environ Microbiol 69:3385–3392
D’Elia MA, Pereira MP, Chung YS et al (2006) Lesions in teichoic acid biosynthesis in Staphylococcus aureus lead to a lethal gain of function in the otherwise dispensable pathway. J Bacteriol 188:4183–4189
Meighen EA (1993) Bacterial bioluminescence: organization, regulation, and application of the lux genes. FASEB J 7:1016–1022
Murray RW, Melchior EP, Hagadorn JC et al (2001) Staphylococcus aureus cell extract transcription-translation assay: firefly luciferase reporter system for evaluating protein translation inhibitors. Antimicrob Agents Chemother 45:1900–1904
Steidler L, Yu W, Fiers W et al (1996) The expression of the Photinus pyralis luciferase gene in Staphylococcus aureus Cowan I allows the development of a live amplifiable tool for immunodetection. Appl Environ Microbiol 62:2356–2359
Meighen EA (1991) Molecular biology of bacterial bioluminescence. Microbiol Rev 55:123–142
Mesak LR, Yim G, Davies J (2009) Improved lux reporters for use in Staphylococcus aureus. Plasmid 61:182–187
Francis KP, Joh D, Bellinger-Kawahara C et al (2000) Monitoring bioluminescent Staphylococcus aureus infections in living mice using a novel luxABCDE construct. Infect Immun 68:3594–3600
Malone CL, Boles BR, Lauderdale KJ et al (2009) Fluorescent reporters for Staphylococcus aureus. J Microbiol Methods 77:251–260
Cheung AL, Nast CC, Bayer AS (1998) Selective activation of sar promoters with the use of green fluorescent protein transcriptional fusions as the detection system in the rabbit endocarditis model. Infect Immun 66:5988–5993
Qazi SN, Rees CE, Mellits KH et al (2001) Development of gfp vectors for expression in Listeria monocytogenes and other low G+C gram positive bacteria. Microb Ecol 41:301–309
Franke GC, Dobinsky S, Mack D et al (2007) Expression and functional characterization of gfpmut3.1 and its unstable variants in Staphylococcus epidermidis. J Microbiol Methods 71:123–132
Andersen JB, Sternberg C, Poulsen LK et al (1998) New unstable variants of green fluorescent protein for studies of transient gene expression in bacteria. Appl Environ Microbiol 64:2240–2246
Pereira PM, Veiga H, Jorge AM et al (2010) Fluorescent reporters for studies of cellular localization of proteins in Staphylococcus aureus. Appl Environ Microbiol 76:4346–4353
Liese J, Rooijakkers SH, van Strijp JA et al (2012) Intravital two-photon microscopy of host-pathogen interactions in a mouse model of Staphylococcus aureus skin abscess formation. Cell Microbiol 15:891–909
Kahl BC, Goulian M, van Wamel W et al (2000) Staphylococcus aureus RN6390 replicates and induces apoptosis in a pulmonary epithelial cell line. Infect Immun 68:5385–5392
Paprotka K, Giese B, Fraunholz MJ (2010) Codon-improved fluorescent proteins in investigation of Staphylococcus aureus host pathogen interactions. J Microbiol Methods 83:82–86
Sastalla I, Chim K, Cheung GY et al (2009) Codon-optimized fluorescent proteins designed for expression in low-GC gram-positive bacteria. Appl Environ Microbiol 75:2099–2110
Pédelacq JD, Cabantous S, Tran T et al (2006) Engineering and characterization of a superfolder green fluorescent protein. Nat Biotechnol 24:79–88
Yu W, Götz F (2012) Cell wall antibiotics provoke accumulation of anchored mCherry in the cross wall of Staphylococcus aureus. PLoS One 7:e30076
Kloos W, Schleifer KH, Götz F (1991) The genus Staphylococcus. In: Balows B, Trüper HG, Dworkin M, Harder W, Schleifer KH (eds) The prokaryotes, vol 2. Springer, London, pp 1369–1420
Halfmann G, Götz F, Lubitz W (1993) Expression of bacteriophage PhiX174 lysis gene E in Staphylococcus carnosus TM300. FEMS Microbiol Lett 108:139–143
Gauger T, Weihs F, Mayer S et al (2012) Intracellular monitoring of target protein production in Staphylococcus aureus by peptide tag-induced reporter fluorescence. Microb Biotechnol 5:129–134
D’Elia MA, Pereira MP, Brown ED (2009) Are essential genes really essential? Trends Microbiol 17:433–438
Sheehan BJ, Foster TJ, Dorman CJ et al (1992) Osmotic and growth-phase dependent regulation of the eta gene of Staphylococcus aureus: a role for DNA supercoiling. Mol Gen Genet 232:49–57
Wang PZ, Projan SJ, Leason KR et al (1987) Translational fusion with a secretory enzyme as an indicator. J Bacteriol 169:3082–3087
Otto M, Süssmuth R, Jung G et al (1998) Structure of the pheromone peptide of the Staphylococcus epidermidis agr system. FEBS Lett 424:89–94
Ohlsen K, Koller KP, Hacker J (1997) Analysis of expression of the alpha-toxin gene (hla) of Staphylococcus aureus by using a chromosomally encoded hla::lacZ gene fusion. Infect Immun 65:3606–3614
Arnaud M, Chastanet A, Debarbouille M (2004) New vector for efficient allelic replacement in naturally nontransformable, low-GC-content, gram-positive bacteria. Appl Environ Microbiol 70:6887–6891
Acknowledgements
Work in the author’s lab was supported by grant BE4038/1 of the Deutsche Forschungsgemeinschaft (DFG).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2014 Springer Science+Business Media New York
About this protocol
Cite this protocol
Bertram, R. (2014). Complementation Plasmids, Inducible Gene-Expression Systems, and Reporters for Staphylococci. In: Bose, J. (eds) The Genetic Manipulation of Staphylococci. Methods in Molecular Biology, vol 1373. Humana Press, New York, NY. https://doi.org/10.1007/7651_2014_181
Download citation
DOI: https://doi.org/10.1007/7651_2014_181
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3157-6
Online ISBN: 978-1-4939-3158-3
eBook Packages: Springer Protocols