Analysis of Polyhydroxyalkanoates Granules in Haloferax mediterranei by Double-Fluorescence Staining with Nile Red and SYBR Green by Confocal Fluorescence Microscopy
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http://hdl.handle.net/10045/115563
Títol: | Analysis of Polyhydroxyalkanoates Granules in Haloferax mediterranei by Double-Fluorescence Staining with Nile Red and SYBR Green by Confocal Fluorescence Microscopy |
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Autors: | Cánovas, Verónica | García-Chumillas, Salvador | Monzó Sánchez, Fuensanta | Simó Cabrera, Lorena | Fernández-Ayuso, Carmen | Pire, Carmen | Martínez-Espinosa, Rosa María |
Grups d'investigació o GITE: | Bioquímica Aplicada/Applied Biochemistry (AppBiochem) |
Centre, Departament o Servei: | Universidad de Alicante. Departamento de Agroquímica y Bioquímica | Universidad de Alicante. Instituto Multidisciplinar para el Estudio del Medio "Ramón Margalef" |
Paraules clau: | Haloarchaea | Haloferax mediterranei | Polyhydroxyalkanoates (PHAs) | Nile red | SYBR Green | Confocal microscopy | Transmission electronic microscopy |
Àrees de coneixement: | Bioquímica y Biología Molecular |
Data de publicació: | 14-de maig-2021 |
Editor: | MDPI |
Citació bibliogràfica: | Cánovas V, Garcia-Chumillas S, Monzó F, Simó-Cabrera L, Fernández-Ayuso C, Pire C, Espinosa RMM. Analysis of Polyhydroxyalkanoates Granules in Haloferax mediterranei by Double-Fluorescence Staining with Nile Red and SYBR Green by Confocal Fluorescence Microscopy. Polymers. 2021; 13(10):1582. https://doi.org/10.3390/polym13101582 |
Resum: | Haloferaxmediterranei is a haloarchaeon of high interest in biotechnology because it produces and mobilizes intracellular polyhydroxyalkanoate (PHA) granules during growth under stress conditions (limitation of phosphorous in the culture media), among other interesting metabolites (enzymes, carotenoids, etc.). The capability of PHA production by microbes can be monitored with the use of staining-based methods. However, the staining of haloarchaea cells is a challenging task; firstly, due to the high ionic strength of the medium, which is inappropriate for most of dyes, and secondly, due to the low permeability of the haloarchaea S-layer to macromolecules. In this work, Haloferax mediterranei is used as a halophilic archaeon model to describe an optimized protocol for the visualization and analysis of intracellular PHA granules in living cells. The method is based on double-fluorescence staining using Nile red and SYBR Green by confocal fluorescence microscopy. Thanks to this method, the capability of PHA production by new haloarchaea isolates could be easily monitored. |
Patrocinadors: | This work was funded by a research grant from Instituto de Fomento de la Region de Murcia (INFO), Spain (2020.08.CT01.0039) and the University of Alicante (VIGROB-309). |
URI: | http://hdl.handle.net/10045/115563 |
ISSN: | 2073-4360 |
DOI: | 10.3390/polym13101582 |
Idioma: | eng |
Tipus: | info:eu-repo/semantics/article |
Drets: | © 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
Revisió científica: | si |
Versió de l'editor: | https://doi.org/10.3390/polym13101582 |
Apareix a la col·lecció: | INV - AppBiochem - Artículos de Revistas |
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Canovas_etal_2021_Polymers.pdf | 3,07 MB | Adobe PDF | Obrir Vista prèvia | |
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