- Protocol for HeLa Cells Infection with Escherichia coli Strains Producing Colibactin and Quantification of the Induced DNA-damage.
Protocol for HeLa Cells Infection with Escherichia coli Strains Producing Colibactin and Quantification of the Induced DNA-damage.
Bio-protocol (2017-08-20)
Nadège Bossuet-Greif, Marcy Belloy, Michèle Boury, Eric Oswald, Jean-Philippe Nougayrede
PMID34541180
ABSTRACT
Strains of Escherichia coli bearing the pks genomic island synthesize the genotoxin colibactin. Exposure of eukaryotic cells to E. coli producing colibactin induces DNA damages, ultimately leading to cell cycle arrest, senescence and death. Here we describe a simple method to demonstrate the genotoxicity of bacteria producing colibactin following a short infection of cultured mammalian cells with pks + E. coli.
MATERIALS
Product Number
Brand
Product Description
Sigma-Aldrich
Dulbecco′s Phosphate Buffered Saline, 10×, Modified, without calcium chloride and magnesium chloride, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Dulbecco′s Phosphate Buffered Saline, Modified, without calcium chloride and magnesium chloride, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Gentamicin solution, 50 mg/mL in deionized water, liquid, 0.1 μm filtered, BioReagent, suitable for cell culture
Sigma-Aldrich
Hanks′ Balanced Salt solution, Modified, with sodium bicarbonate, without phenol red, liquid, sterile-filtered, suitable for cell culture