Abstract
The ability to create mutations is an important step towards understanding bacterial physiology and virulence. While targeted approaches are invaluable, the ability to produce genome-wide random mutations can lead to crucial discoveries. Transposon mutagenesis is a useful approach, but many interesting mutations can be missed by these insertions that interrupt coding and noncoding sequences due to the integration of an entire transposon. Chemical mutagenesis and UV-based random mutagenesis are alternate approaches to isolate mutations of interest with the potential of only single nucleotide changes. Once a standard method, difficulty in identifying mutation sites had decreased the popularity of this technique. However, thanks to the recent emergence of economical whole-genome sequencing, this approach to making mutations can once again become a viable option. Therefore, this chapter provides an overview protocol for random mutagenesis using UV light or DNA-damaging chemicals.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Ikehata H, Ono T (2011) The mechanisms of UV mutagenesis. J Radiat Res 52:115–125
Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory, New York, NY
Barnes IJ, Bondi A, Moat AG (1969) Biochemical characterization of lysine auxotrophs of Staphylococcus aureus. J Bacteriol 99:169–174
Dornbusch K, Hallander HO, Lofquist F (1969) Extrachromosomal control of methicillin resistance and toxin production in Staphylococcus aureus. J Bacteriol 98:351–358
Goering RV, Pattee PA (1971) Mutants of Staphylococcus aureus with increased sensitivity to ultraviolet radiation. J Bacteriol 106:157–161
Markham PN, Westhaus E, Klyachko K et al (1999) Multiple novel inhibitors of the NorA multidrug transporter of Staphylococcus aureus. Antimicrob Agents Chemother 43:2404–2408
Matthews PR, Reed KC, Stewart PR (1987) The cloning of chromosomal DNA associated with methicillin and other resistances in Staphylococcus aureus. J Gen Microbiol 133:1919–1929
Silverman JA, Oliver N, Andrew T et al (2001) Resistance studies with daptomycin. Antimicrob Agents Chemother 45:1799–1802
Kreiswirth BN, Lofdahl S, Betley MJ et al (1983) The toxic shock syndrome exotoxin structural gene is not detectably transmitted by a prophage. Nature 305:709–712
Ronen A (1964) Back mutation of leucine-requiring auxotrophs of Salmonella Typhimurium induced by diethylsulphate. J Gen Microbiol 37:49–58
Adelberg EA, Mandel M, Chen GCC (1965) Optimal conditions for mutagenesis by N-methyl-N′-nitro-N-nitrosoguanidine in Escherichia coli K12. Biochem Biophys Res Commun 18:788–795
Chapple RM, Inglis B, Stewart PR (1992) Lethal and mutational effects of solar and UV radiation on Staphylococcus aureus. Arch Microbiol 157:242–248
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
Bose, J.L. (2014). Chemical and UV Mutagenesis. 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_190
Download citation
DOI: https://doi.org/10.1007/7651_2014_190
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