Benincasa Monica, Barrière Quentin, Runti Giulia, Pierre Olivier, Bourge Mick, Scocchi Marco, Mergaert Peter
Department of Life Sciences, University of Trieste, Trieste, Italy.
Institute for Integrative Biology of the Cell (I2BC), CEA, CNRS, University Paris-Sud, Université Paris-Saclay, Gif-sur-Yvette cedex, France.
Bio Protoc. 2016 Dec 5;6(23):e2038. doi: 10.21769/BioProtoc.2038.
Antimicrobial peptides (AMPs) can target the bacterial envelope or alternatively have intracellular targets. The latter requires uptake of the peptide by the bacterial cells. The bacterial internalization of an AMP can be evaluated by a fluorescence-based method that couples the use of the fluorescently labelled AMP to the fluorescence quencher trypan blue. Trypan blue is excluded from the interior of intact cells and the fluorescence of the extracellular peptide or of the peptide bound on the bacterial surface can be quenched by it, while the fluorescence of the internalized peptide is not affected. The uptake of the peptide by the bacteria is determined by measuring the fluorescence in individual cells by flow cytometry.
抗菌肽（AMPs）可以靶向细菌包膜，或者具有细胞内靶点。后者要求肽被细菌细胞摄取。抗菌肽的细菌内化作用可以通过一种基于荧光的方法来评估，该方法将荧光标记的抗菌肽与荧光淬灭剂台盼蓝结合使用。完整细胞内部可排除台盼蓝，细胞外肽或结合在细菌表面的肽的荧光可被其淬灭，而内化肽的荧光不受影响。通过流式细胞术测量单个细胞中的荧光来确定细菌对肽的摄取。
