Expression of NanoLuc Luciferase in Listeria innocua for Development of Biofilm Assay

Autor: Aleš Berlec, Nika Janež, Meta Sterniša, Anja Klančnik, Jerica Sabotič
Jazyk: angličtina
Rok vydání: 2021
Předmět:
Zdroj: Frontiers in Microbiology, Vol 12 (2021)
Druh dokumentu: article
ISSN: 1664-302X
DOI: 10.3389/fmicb.2021.636421
Popis: Studies of biofilm formation by bacteria are crucial for understanding bacterial resistance and for development of novel antibacterial strategies. We have developed a new bioluminescence biofilm assay for Listeria innocua, which is considered a non-pathogenic surrogate for Listeria monocytogenes. L. innocua was transformed with a plasmid for inducible expression of NanoLuc luciferase (Nluc). Concentration-dependent bioluminescence signals were obtained over a concentration range of more than three log units. This biofilm assay enables absolute quantification of bacterial cells, with the necessary validation. For biofilm detection and quantification, this “Nluc bioluminescence” method has sensitivity of 1.0 × 104 and 3.0 × 104 colony forming units (CFU)/mL, respectively, with a dynamic range of 1.0 × 104 to 5.0 × 107 CFU/mL. These are accompanied by good precision (coefficient of variation,
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