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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| Format: | Article |
| Language: | English |
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Wiley
2017-01-01
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| Series: | Journal of Parasitology Research |
| Online Access: | http://dx.doi.org/10.1155/2017/1964531 |
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| author | Andrés Vacas Conor Sugden Óscar Velasco-Rodriguez Miriam Algarabel-Olona José Peña-Guerrero Esther Larrea Celia Fernández-Rubio Paul A. Nguewa |
| author_facet | Andrés Vacas Conor Sugden Óscar Velasco-Rodriguez Miriam Algarabel-Olona José Peña-Guerrero Esther Larrea Celia Fernández-Rubio Paul A. Nguewa |
| author_sort | Andrés Vacas |
| collection | DOAJ |
| 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. |
| format | Article |
| id | doaj-art-3a1af7eba0bf4a91b3c10c7f812e5da8 |
| institution | Kabale University |
| issn | 2090-0023 2090-0031 |
| language | English |
| publishDate | 2017-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of Parasitology Research |
| spelling | doaj-art-3a1af7eba0bf4a91b3c10c7f812e5da82025-02-03T06:11:16ZengWileyJournal of Parasitology Research2090-00232090-00312017-01-01201710.1155/2017/19645311964531Construction of Two mCherry Plasmids (pXG-mCherry) for Transgenic Leishmania: Valuable Tools for Future Molecular AnalysisAndrés Vacas0Conor Sugden1Óscar Velasco-Rodriguez2Miriam Algarabel-Olona3José Peña-Guerrero4Esther Larrea5Celia Fernández-Rubio6Paul A. Nguewa7Instituto de Salud Tropical (ISTUN), University of Navarra, Pamplona, SpainInstituto de Salud Tropical (ISTUN), University of Navarra, Pamplona, SpainInstituto de Salud Tropical (ISTUN), University of Navarra, Pamplona, SpainInstituto de Salud Tropical (ISTUN), University of Navarra, Pamplona, SpainInstituto de Salud Tropical (ISTUN), University of Navarra, Pamplona, SpainInstituto de Salud Tropical (ISTUN), University of Navarra, Pamplona, SpainInstituto de Salud Tropical (ISTUN), University of Navarra, Pamplona, SpainInstituto de Salud Tropical (ISTUN), University of Navarra, Pamplona, SpainLeishmania 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.http://dx.doi.org/10.1155/2017/1964531 |
| spellingShingle | Andrés Vacas Conor Sugden Óscar Velasco-Rodriguez Miriam Algarabel-Olona José Peña-Guerrero Esther Larrea Celia Fernández-Rubio Paul A. Nguewa Construction of Two mCherry Plasmids (pXG-mCherry) for Transgenic Leishmania: Valuable Tools for Future Molecular Analysis Journal of Parasitology Research |
| 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 |
| url | http://dx.doi.org/10.1155/2017/1964531 |
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