Abstract
Bacterial luminescence allows for noninvasive continuous monitoring of promoter activity in a wide range of model systems. This chapter details various examples of use of the lux reporter system to measure promoter activity in bacteria using the vector pUC18T-mini-Tn7T-lux-Tp. Here, we describe the construction of promoter fusions with bacterial luciferase, and how to quantify promoter activity in real time in vitro and in vivo in plant, insect, and murine infection models.
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References
Baldwin TO, Christopher JA, Raushel FM et al (1995) Structure of bacterial luciferase. Curr Opin Struct Biol 5:798–809
Choi K-H, Schweizer HP (2006) Mini-Tn7 insertion in bacteria with single attTn7 sites: example Pseudomonas aeruginosa. Nat Protoc 1:153–161. https://doi.org/10.1038/nprot.2006.24
Choi K-H, Gaynor JB, White KG et al (2005) A Tn7-based broad-range bacterial cloning and expression system. Nat Methods 2:443–448. https://doi.org/10.1038/nmeth765
Damron FH, McKenney ES, Barbier M et al (2013) Construction of mobilizable mini-Tn7 vectors for bioluminescent detection of gram-negative bacteria and single-copy promoter lux reporter analysis. Appl Environ Microbiol 79:4149–4153. https://doi.org/10.1128/AEM.00640-13
Choi K-H, Mima T, Casart Y et al (2008) Genetic tools for select-agent-compliant manipulation of Burkholderia pseudomallei. Appl Environ Microbiol 74:1064–1075. https://doi.org/10.1128/AEM.02430-07
Reed LJ, Muench H (1938) A simple method of estimating fifty per cent endpoints. Am J Epidemiol 27:493–497
Schneider CA, Rasband WS, Eliceiri KW (2012) NIH image to ImageJ: 25 years of image analysis. Nat Methods 9:671–675
Weiner ZP, Ernst SM, Boyer AE et al (2014) Circulating lethal toxin decreases the ability of neutrophils to respond to Bacillus anthracis. Cell Microbiol 16:504–518. https://doi.org/10.1111/cmi.12232
Choi KH, Kumar A, Schweizer HP (2006) A 10-min method for preparation of highly electrocompetent Pseudomonas aeruginosa cells: application for DNA fragment transfer between chromosomes and plasmid transformation. J Microbiol Methods 64:391–397. https://doi.org/10.1016/j.mimet.2005.06.001
Sambrook DWJR (2001) Molecular cloning: a laboratory manual, 3rd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor
Figurski DH, Helinski DR (1979) Replication of an origin-containing derivative of plasmid RK2 dependent on a plasmid function provided in trans. Proc Natl Acad Sci U S A 76:1648–1652
López CM, Rholl DA, Trunck LA, Schweizer HP (2009) Versatile dual-technology system for markerless allele replacement in Burkholderia pseudomallei. Appl Environ Microbiol 75:6496–6503. https://doi.org/10.1128/AEM.01669-09
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Barbier, M., Bevere, J., Damron, F.H. (2018). In Vivo Bacterial Imaging Using Bioluminescence. In: Dubey, P. (eds) Reporter Gene Imaging. Methods in Molecular Biology, vol 1790. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7860-1_7
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DOI: https://doi.org/10.1007/978-1-4939-7860-1_7
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