Abstract
Listeria monocytogenes is a Gram-positive, facultative intracellular foodborne pathogen that enters the human digestive tract after the consumption of contaminated food. Much research has been done to understand the virulence factors of Listeria monocytogenes. One useful tool to study these virulence factors has been transposon mutagenesis. Many mutants can be generated at a time by performing high-throughput mutagenesis using transposons and later screening these mutants to identify features related to particular functions in the bacteria. Many transposon delivery systems are not ideal for transposon studies in Listeria monocytogenes, as the transposon system is too large, has lower transposition efficiency, and a high rate of plasmid retention. Therefore, a new mariner-based transposition system has been developed for Listeria monocytogenes. This system is an ideal high-throughput transposon mutagenesis as the rate of transposition is high and random, along with very low plasmid retention capacity.
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References
Ryser ET, Marth EH (1989) “New” food-borne pathogens of public health significance. J Am Diet Assoc 89:948–954
Farber JM, Peterkin PI (1991) Listeria monocytogenes, a food-borne pathogen. Microbiol Rev 55:476–511
Kwon YM, Ricke SC, Mandal RK (2016) Transposon sequencing: methods and expanding applications. Appl Microbiol Biotechnol 100:31–43
Annous BA, Becker LA, Bayles DO, Labeda DP, Wilkinson BJ (1997) Critical role of anteiso-C15:0 fatty acid in the growth of Listeria monocytogenes at low temperatures. Appl Environ Microbiol 63:3887–3894
Zhu K, Bayles DO, Xiong A, Jayaswal RK, Wilkinson BJ (2005) Precursor and temperature modulation of fatty acid composition and growth of Listeria monocytogenes cold-sensitive mutants with transposon-interrupted branched-chain alpha-keto acid dehydrogenase. Microbiology 151:615–623
Angelidis AS, Smith LT, Hoffman LM, Smith GM (2002) Identification of opuC as a chill-activated and osmotically activated carnitine transporter in Listeria monocytogenes. Appl Environ Microbiol 68:2644–2650
Sleator RD, Wouters J, Gahan CG, Abee T, Hill C (2001) Analysis of the role of OpuC, an osmolyte transport system, in salt tolerance and virulence potential of Listeria monocytogenes. Appl Environ Microbiol 67:2692–2698
Joseph B, Przybilla K, Stuhler C, Schauer K, Slaghuis J, Fuchs TM, Goebel W (2006) Identification of Listeria monocytogenes genes contributing to intracellular replication by expression profiling and mutant screening. J Bacteriol 188:556–568
Gaillard JL, Berche P, Sansonetti P (1986) Transposon mutagenesis as a tool to study the role of hemolysin in the virulence of Listeria monocytogenes. Infect Immun 52:50–55
Collatz E, Carlier C, Courvalin P (1984) Characterization of high-level aminoglycoside resistance in a strain of Streptococcus pneumoniae. J Gen Microbiol 130:1665–1671
Autret N, Dubail I, Trieu-Cuot P, Berche P, Charbit A (2001) Identification of new genes involved in the virulence of Listeria monocytogenes by signature-tagged transposon mutagenesis. Infect Immun 69:2054–2065
Kathariou S, Metz P, Hof H, Goebel W (1987) Tn916-induced mutations in the hemolysin determinant affecting virulence of Listeria monocytogenes. J Bacteriol 169:1291–1297
Camilli A, Portnoy A, Youngman P (1990) Insertional mutagenesis of Listeria monocytogenes with a novel Tn917 derivative that allows direct cloning of DNA flanking transposon insertions. J Bacteriol 172:3738–3744
Muller A, Rychli K, Muhterem-Uyar M, Zaiser A, Stessl B, Guinane CM, Cotter PD, Wagner M, Schmitz-Esser S (2013) Tn6188 - a novel transposon in Listeria monocytogenes responsible for tolerance to benzalkonium chloride. PLoS One 8:e76835
Muller A, Rychli K, Zaiser A, Wieser C, Wagner M, Schmitz-Esser S (2014) The Listeria monocytogenes transposon Tn6188 provides increased tolerance to various quaternary ammonium compounds and ethidium bromide. FEMS Microbiol Lett 361:166–173
Cummins J, Casey PG, Joyce SA, Gahan CG (2013) A mariner transposon-based signature-tagged mutagenesis system for the analysis of oral infection by Listeria monocytogenes. PLoS One 8:e75437. https://doi.org/10.1371/journal.pone.0075437
Picardeau M (2010) Transposition of fly mariner elements into bacteria as a genetic tool for mutagenesis. Genetica 138:551–558
Plasterk RH, Izsvak Z, Ivics Z (1999) Resident aliens: the Tc1/mariner superfamily of transposable elements. Trends Genet 15:326–332
Lampe DJ, Churchill ME, Robertson HM (1996) A purified mariner transposase is sufficient to mediate transposition in vitro. EMBO J 15:5470–5479
Cao M, Bitar AP, Marquis H (2007) A mariner-based transposition system for Listeria monocytogenes. Appl Environ Microbiol 73:2758–2761
Garsin DA, Urbach J, Huguet-Tapia JC, Peters JE, Ausubel FM (2004) Construction of an Enterococcus faecalis Tn917-mediated-gene-disruption library offers insight into Tn917 insertion patterns. J Bacteriol 186:7280–7289
Zemansky J, Kline BC, Woodward JJ, Leber JH, Marquis H, Portnoy DA (2009) Development of a mariner-based transposon and identification of Listeria monocytogenes determinants, including the peptidyl-prolyl isomerase PrsA2, that contribute to its hemolytic phenotype. J Bacteriol 191:3950–3964
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Paul, O., Chakravarty, D., Donaldson, J.R. (2019). Transposon Mutagenesis of Listeria monocytogenes . In: Ricke, S., Park, S., Davis, M. (eds) Microbial Transposon Mutagenesis. Methods in Molecular Biology, vol 2016. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9570-7_6
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DOI: https://doi.org/10.1007/978-1-4939-9570-7_6
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