The authors have declared no competing interest. Together, our results show that BaCAPA can effectively report on the accumulation of molecules into bacterial cells and we anticipate that this will widen the set of tools available to generally measure permeability into bacterial cells. Gram-negative bacilli are a large group of bacteria that are commonly found in the intestinal tract of humans and most animals. Significantly, we also showed that BaCAPA can be used to analyze the permeation of molecules within the phagocytes of macrophages, which may prove critical in the analysis of intracellular pathogens. coli) enabled a compartmental analysis of permeation across individual cell membranes. Directing the localization of the HaloTag protein to either the cytoplasm or periplasm of Escherichia coli ( E. Herein, we describe the development of the Bacterial Chloro- Alkane Penetration Assay (BaCAPA), which employs the use of a genetically encoded protein called HaloTag, to measure the uptake and accumulation of molecules into Gram-negative bacteria.
While it is well appreciated that the OM is a principal determinant of small molecule permeation, it has proven to be challenging to assess this feature in a robust and quantitative way. aeruginosa en Acinetobacter spp in Belgische laboratoria. Algemene richtlijnen voor de detectie van Carbapenemases bij multi-resistente P. For Gram-negative bacteria, this second exterior membrane is known as the Outer Membrane (OM) and its unique composition is generally what restricts the passive permeation of small molecules into bacterial cells. Algemene richtlijnen voor detectie van CPE in laboratoria in België. 1 They are characterized by their cell envelopes, which are composed of a thin peptidoglycan cell wall sandwiched between an inner cytoplasmic cell membrane and a bacterial outer membrane. Some of the most dangerous bacterial pathogens (Gram-negative and mycobacteria) deploy a formidable secondary membrane barrier to reduce the influx of exogenous molecules. Gram-negative bacteria are bacteria that do not retain the crystal violet stain used in the Gram staining method of bacterial differentiation.
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