Abstract
Pathogenic bacteria can have sub-populations of hypermutable bacteria. This sub-population has a higher spontaneous mutation rate than the majority of the population which can be attributed to defects in proofreading and repair mechanisms. This leads to the evolution of drug-resistant strains of bacteria through genetic change. It is important to study the expression of genes involved in, for example, mismatch repair and the SOS system by real-time PCR to determine hypermutability and therefore provide an indicator of the mutagenic ability of certain strains of pathogenic bacteria.
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References
Miller JH (1996) Spontaneous mutators in bacteria: insights into pathways of mutagenesis and repair. Annu Rev Microbiol 50:625–643
Oliver A, Baquero F, Blazquez J (2002) The mismatch repair system (mutS, mutL and uvrD genes) in Pseudomonas aeruginosa: molecular characterization of naturally occurring mutants. Mol Microbiol 43:1641–1650
Horst J-P, Wu T-H, Marinus MG (1999) Escherichia coli mutator genes. Trends Microbiol 7:29–36
Herman GE, Modrich P (1981) Escherichia coli K-12 clones that overproduce dam methylase are hypermutable. J Bacteriol 145:644–646
Oliver A, Canton R, Campo P, Baquero F, Blazquez J (2000) High frequency of hypermutable Pseudomonas aeruginosa in cystic fibrosis lung infection. Science 288:1251–1254
Wilson M, DeRisi J, Kristensen H-H, Imboden P, Rane S, Brown PO, Schoolnik GK (1999) Exploring drug-induced alterations in gene expression in Mycobacterium tuberculosis by microarray hybridization. Proc Natl Acad Sci USA 96:12833–12838
Doumith M, Cazalet C, Simoes N, Frangeul L, Jacquet C, Kunst F, Martin P, Cossart P, Glaser P, Buchrieser C (2004) New aspects regarding evolution and virulence of Listeria monocytogenes revealed by comparative genomics and DNA arrays. Infect Immun 72:1072–1083
Saunders NA, Underwood A, Kearns AM, Hallas G (2004) A virulence-associated gene microarray: a tool for investigation of the evolution and pathogenic potential of Staphylococcus aureus. Microbiology 150:3763–3771
Marton MJ, DeRisi JL, Bennett HA, Iyer VR, Meyer MR, Roberts CJ, Stoughton R, Burchard J, Slade D, Dai H, Bassett DE, Hartwell LH, Brown PO, Friend SH (1998) Drug target validation and identification of secondary drug target effects using DNA microarrays. Nat Med 4:1293–1301
Adam M, Murali B, Glenn N, Potter SS (2008) Epigenetic inheritance based evolution of antibiotic resistance in bacteria. BMC Evol Biol 8:52
Guard-Bouldin J, Morales CA, Frye JG, Gast RK, Musgrove M (2007) Detection of Salmonella enterica subpopulations by phenotype microarray antibiotic resistance patterns. Appl Environ Microbiol 73:7753–7756
Dumas JL, van Delden C, Perron K, Köhler T (2006) Analysis of antibiotic resistance gene expression in Pseudomonas aeruginosa by quantitative real-time-PCR. FEMS Microbiol Lett 254:217–225
Mangan JA, Monahan IM, Butcher PD (2002) Gene expression during host-pathogen interactions: approaches to bacterial mRNA extraction and labelling for microarray analysis. In: Wren B, Dorrell N (eds) Functional microbial genomics: methods in microbiology. Academic, London, pp 137–151
Brooks PC, Movahedzadeh F, Davis EO (2001) Identification of some DNA damage-inducible genes of Mycobacterium tuberculosis: apparent lack of correlation with LexA binding. J Bacteriol 183:4459–4467
Boshoff HI, Reed MB, Barry CE III, Mizrahi V (2003) DnaE2 polymerase contributes to in vivo survival and the emergence of drug resistance in Mycobacterium tuberculosis. Cell 113:183–193
Hampshire T, Soneji S, Bacon J, James BW, Hinds J, Laing K, Stabler RA, Marsh PD, Butcher PD (2004) Stationary phase gene expression of Mycobacterium tuberculosis following a progressive nutrient depletion: a model for persistent organisms? Tuberculosis 84:228–238
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods 25:402–408
Pfaffl MW (2001) A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res 29:e45
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O’Sullivan, D.M. (2010). Real-Time PCR Methods to Study Expression of Genes Related to Hypermutability. In: Gillespie, S., McHugh, T. (eds) Antibiotic Resistance Protocols. Methods in Molecular Biology, vol 642. Humana Press. https://doi.org/10.1007/978-1-60327-279-7_5
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DOI: https://doi.org/10.1007/978-1-60327-279-7_5
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