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Construction of Two mCherry Plasmids (pXG-mCherry) for Transgenic Leishmania: Valuable Tools for Future Molecular Analysis

Leishmania is the causative agent of leishmaniasis, a neglected tropical disease that affects more than 12 million people around the world. Current treatments are toxic and poorly effective due to the acquisition of resistance within Leishmania populations. Thus, the pursuit for new antileishmanial...

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Autores principales: Vacas, Andrés, Sugden, Conor, Velasco-Rodriguez, Óscar, Algarabel-Olona, Miriam, Peña-Guerrero, José, Larrea, Esther, Fernández-Rubio, Celia, Nguewa, Paul A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Hindawi Publishing Corporation 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5327783/
https://www.ncbi.nlm.nih.gov/pubmed/28286673
http://dx.doi.org/10.1155/2017/1964531
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author Vacas, Andrés
Sugden, Conor
Velasco-Rodriguez, Óscar
Algarabel-Olona, Miriam
Peña-Guerrero, José
Larrea, Esther
Fernández-Rubio, Celia
Nguewa, Paul A.
author_facet Vacas, Andrés
Sugden, Conor
Velasco-Rodriguez, Óscar
Algarabel-Olona, Miriam
Peña-Guerrero, José
Larrea, Esther
Fernández-Rubio, Celia
Nguewa, Paul A.
author_sort Vacas, Andrés
collection PubMed
description Leishmania is the causative agent of leishmaniasis, a neglected tropical disease that affects more than 12 million people around the world. Current treatments are toxic and poorly effective due to the acquisition of resistance within Leishmania populations. Thus, the pursuit for new antileishmanial drugs is a priority. The available methods for drug screening based on colorimetric assays using vital dyes are time-consuming. Currently, the use of fluorescent reporter proteins is replacing the use of viability indicator dyes. We have constructed two plasmids expressing the red fluorescent protein mCherry with multiple cloning sites (MCS), adequate for N- and C-terminal fusion protein constructs. Our results also show that the improved pXG-mCherry plasmid can be employed for drug screening in vitro. The use of the red fluorescent protein, mCherry, is an easier tool for numerous assays, not only to test pharmacological compounds, but also to determine the subcellular localization of proteins.
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spelling pubmed-53277832017-03-12 Construction of Two mCherry Plasmids (pXG-mCherry) for Transgenic Leishmania: Valuable Tools for Future Molecular Analysis Vacas, Andrés Sugden, Conor Velasco-Rodriguez, Óscar Algarabel-Olona, Miriam Peña-Guerrero, José Larrea, Esther Fernández-Rubio, Celia Nguewa, Paul A. J Parasitol Res Research Article Leishmania is the causative agent of leishmaniasis, a neglected tropical disease that affects more than 12 million people around the world. Current treatments are toxic and poorly effective due to the acquisition of resistance within Leishmania populations. Thus, the pursuit for new antileishmanial drugs is a priority. The available methods for drug screening based on colorimetric assays using vital dyes are time-consuming. Currently, the use of fluorescent reporter proteins is replacing the use of viability indicator dyes. We have constructed two plasmids expressing the red fluorescent protein mCherry with multiple cloning sites (MCS), adequate for N- and C-terminal fusion protein constructs. Our results also show that the improved pXG-mCherry plasmid can be employed for drug screening in vitro. The use of the red fluorescent protein, mCherry, is an easier tool for numerous assays, not only to test pharmacological compounds, but also to determine the subcellular localization of proteins. Hindawi Publishing Corporation 2017 2017-02-13 /pmc/articles/PMC5327783/ /pubmed/28286673 http://dx.doi.org/10.1155/2017/1964531 Text en Copyright © 2017 Andrés Vacas et al. https://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Vacas, Andrés
Sugden, Conor
Velasco-Rodriguez, Óscar
Algarabel-Olona, Miriam
Peña-Guerrero, José
Larrea, Esther
Fernández-Rubio, Celia
Nguewa, Paul A.
Construction of Two mCherry Plasmids (pXG-mCherry) for Transgenic Leishmania: Valuable Tools for Future Molecular Analysis
title Construction of Two mCherry Plasmids (pXG-mCherry) for Transgenic Leishmania: Valuable Tools for Future Molecular Analysis
title_full Construction of Two mCherry Plasmids (pXG-mCherry) for Transgenic Leishmania: Valuable Tools for Future Molecular Analysis
title_fullStr Construction of Two mCherry Plasmids (pXG-mCherry) for Transgenic Leishmania: Valuable Tools for Future Molecular Analysis
title_full_unstemmed Construction of Two mCherry Plasmids (pXG-mCherry) for Transgenic Leishmania: Valuable Tools for Future Molecular Analysis
title_short Construction of Two mCherry Plasmids (pXG-mCherry) for Transgenic Leishmania: Valuable Tools for Future Molecular Analysis
title_sort construction of two mcherry plasmids (pxg-mcherry) for transgenic leishmania: valuable tools for future molecular analysis
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5327783/
https://www.ncbi.nlm.nih.gov/pubmed/28286673
http://dx.doi.org/10.1155/2017/1964531
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