RT Book, Section
T1 Dual-Fluorescence Chromosome-Located Labeling System for Accurate In Vivo Single-Cell Gene Expression Analysis in Pseudomonas syringae.
A1 López-Pagán, Nieves
A1 Rufián, José S.
A1 Ruiz-Albert, Francisco Javier
A1 Beuzón-López, Carmen del Rosario
K1 Genética
K1 Mutación (Biología)
K1 Microscopía de fluorescencia
K1 Citometría de flujo
AB Epigenetic regulation as a means for bacterial adaptation is receiving increasing interest in the last decade. Significant efforts have been directed towards understanding the mechanisms giving raise to phenotypic heterogeneity within bacterial populations and its adaptive relevance. Phenotypic heterogeneity mostly refers to phenotypic variation not linked to genetic differences nor to environmental stimuli. Recent findings on the relevance of phenotypic heterogeneity on some bacterial complex traits are causing a shift from traditional assays where bacterial phenotypes are defined by averaging population-level data, to single-cell analysis that focus on bacterial individual behavior within the population. Fluorescent labeling is a key asset for single-cell gene expression analysis using flow cytometry, fluorescence microscopy, and/or microfluidics.We previously described the generation of chromosome-located transcriptional gene fusions to fluores-cent reporter genes using the model bacterial plant pathogen Pseudomonas syringae. These fusions allow researchers to follow variation in expression of the gene(s) of interest, without affecting gene function. In this report, we improve the analytic power of the method by combining such transcriptional fusions with constitutively expressed compatible fluorescent reporter genes integrated in a second, neutral locus of the bacterial chromosome. Constitutively expressed fluorescent reporters allow for the detection of all bacteria comprising a heterogeneous population, regardless of the level of expression of the concurrently monitored gene of interest, thus avoiding the traditional use of stains often incompatible with samples from complex contexts such as the leaf.
PB Humana Press (Springer Group)
YR 2024
FD 2024-01-25
LK https://hdl.handle.net/10630/37408
UL https://hdl.handle.net/10630/37408
LA eng
NO López-Pagán N, Rufián JS, Ruiz-Albert J, Beuzón CR. Dual-Fluorescence Chromosome-Located Labeling System for Accurate In Vivo Single-Cell Gene Expression Analysis in Pseudomonas syringae. Methods Mol Biol. 2024;2751:95-114
NO https://www.springernature.com/gp/open-science/policies/book-policies
NO Grants RTI2018-095069-B-I00 and PID2021-127245OB-100 funded by MCIN/AEI/10.13039/ 501100011033/ and by “ERDF A way of making Europe”Grant PY18-2398 funded by Plan Andaluz de Investigación, Desarrollo e Innovación (PAIDI 2020)Plan Andaluz de Investigación, Desarrollo e Innovación (PAIDI 2020)
DS RIUMA. Repositorio Institucional de la Universidad de Málaga
RD 20 ene 2026