Red-Shifted Fluorogenic Substrate for Detection of lacZ-Positive Cells in Living Tissue with Single-Cell Resolution.
Angew Chem Int Ed Engl
; 57(48): 15702-15706, 2018 11 26.
Article
en En
| MEDLINE
| ID: mdl-30255610
ABSTRACT
The Escherichia coli lacZ gene encoding ß-galactosidase is a widely used reporter, but few synthetic substrates are available for detecting its activity with single-cell resolution in living samples. Our recently reported fluorogenic substrate SPiDER-ßGal is suitable for this purpose, but its hydrolysis product shows green fluorescence emission, and a red-shifted analogue is therefore required for use in combination with green fluorescent protein (GFP) markers. Herein, we describe the development of a red-shifted fluorogenic substrate for ß-galactosidase, SPiDER-Red-ßGal, based on a silicon rhodol scaffold and a carboxylic group as the intramolecular nucleophile. LacZ-positive cells were successfully labeled with SPiDER-Red-ßGal at single-cell resolution in living samples, which enabled us to visualize different cell types in combination with GFP markers.
Palabras clave
Texto completo:
1
Banco de datos:
MEDLINE
Asunto principal:
Beta-Galactosidasa
/
Escherichia coli
/
Análisis de la Célula Individual
/
Colorantes Fluorescentes
/
Operón Lac
Tipo de estudio:
Diagnostic_studies
Idioma:
En
Año:
2018
Tipo del documento:
Article