- 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
摘要
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.
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Sigma-Aldrich
杜氏磷酸盐缓冲盐水, Modified, without calcium chloride and magnesium chloride, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
杜氏磷酸盐缓冲盐水, 10×, Modified, without calcium chloride and magnesium chloride, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
庆大霉素 溶液, 50 mg/mL in deionized water, liquid, 0.1 μm filtered, BioReagent, suitable for cell culture
Sigma-Aldrich
汉克斯平衡盐 溶液, Modified, with sodium bicarbonate, without phenol red, liquid, sterile-filtered, suitable for cell culture