Development of a novel set of Gateway-compatible vectors for live imaging in insect cells
Insect genomics is a growing area of research. To exploit fully the genomic data that are being generated, high-throughput systems for the functional characterization of insect proteins and their interactomes are required. In this work, a Gateway-compatible vector set for expression of fluorescent f...
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| Format: | info:ar-repo/semantics/artículo |
| Language: | Inglés |
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The Royal Entomological Society
2023
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| Online Access: | http://hdl.handle.net/20.500.12123/15186 https://resjournals.onlinelibrary.wiley.com/doi/10.1111/j.1365-2583.2011.01100.x https://doi.org/10.1111/j.1365-2583.2011.01100.x |
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| author | Maroniche, Guillermo Andrés Mongelli, Vanesa Claudia Alfonso, Victoria Llauger, Gabriela Taboga, Oscar Alberto Del Vas, Mariana |
| author_browse | Alfonso, Victoria Del Vas, Mariana Llauger, Gabriela Maroniche, Guillermo Andrés Mongelli, Vanesa Claudia Taboga, Oscar Alberto |
| author_facet | Maroniche, Guillermo Andrés Mongelli, Vanesa Claudia Alfonso, Victoria Llauger, Gabriela Taboga, Oscar Alberto Del Vas, Mariana |
| author_sort | Maroniche, Guillermo Andrés |
| collection | INTA Digital |
| description | Insect genomics is a growing area of research. To exploit fully the genomic data that are being generated, high-throughput systems for the functional characterization of insect proteins and their interactomes are required. In this work, a Gateway-compatible vector set for expression of fluorescent fusion proteins in insect cells was developed. The vector set was designed to express a protein of interest fused to any of four different fluorescent proteins [green fluorescent protein (GFP), cyan fluorescent protein (CFP), yellow fluorescent protein (YFP) and mCherry] by either the C-terminal or the N-terminal ends. Additionally, a collection of organelle-specific fluorescent markers was assembled for colocalization with fluorescent recombinant proteins of interest. Moreover, the vector set was proven to be suitable for simultaneously detecting up to three proteins by multiple labelling. The use of the vector set was exemplified by defining the subcellular distribution of Mal de Río Cuarto virus (MRCV) outer coat protein P10 and by analysing the in vivo self-interaction of the MRCV viroplasm matrix protein P9-1 in Förster resonance energy transfer (FRET) experiments. In conclusion, we have developed a valuable tool for high-throughput studies of protein subcellular localization that will aid in the elucidation of the function of newly described insect and virus proteins. |
| format | info:ar-repo/semantics/artículo |
| id | INTA15186 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2023 |
| publishDateRange | 2023 |
| publishDateSort | 2023 |
| publisher | The Royal Entomological Society |
| publisherStr | The Royal Entomological Society |
| record_format | dspace |
| spelling | INTA151862023-09-12T14:01:41Z Development of a novel set of Gateway-compatible vectors for live imaging in insect cells Maroniche, Guillermo Andrés Mongelli, Vanesa Claudia Alfonso, Victoria Llauger, Gabriela Taboga, Oscar Alberto Del Vas, Mariana Vectors Genomics Insects Viruses Vectores Genómica Insectos Virus Mal de Río Cuarto virus Virus del Mal de Río Cuarto Insect genomics is a growing area of research. To exploit fully the genomic data that are being generated, high-throughput systems for the functional characterization of insect proteins and their interactomes are required. In this work, a Gateway-compatible vector set for expression of fluorescent fusion proteins in insect cells was developed. The vector set was designed to express a protein of interest fused to any of four different fluorescent proteins [green fluorescent protein (GFP), cyan fluorescent protein (CFP), yellow fluorescent protein (YFP) and mCherry] by either the C-terminal or the N-terminal ends. Additionally, a collection of organelle-specific fluorescent markers was assembled for colocalization with fluorescent recombinant proteins of interest. Moreover, the vector set was proven to be suitable for simultaneously detecting up to three proteins by multiple labelling. The use of the vector set was exemplified by defining the subcellular distribution of Mal de Río Cuarto virus (MRCV) outer coat protein P10 and by analysing the in vivo self-interaction of the MRCV viroplasm matrix protein P9-1 in Förster resonance energy transfer (FRET) experiments. In conclusion, we have developed a valuable tool for high-throughput studies of protein subcellular localization that will aid in the elucidation of the function of newly described insect and virus proteins. Instituto de Biotecnología Fil: Maroniche, Guillermo Andrés. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina Fil: Mongelli, Vanesa Claudia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina Fil: Alfonso, Victoria. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina Fil: Llauger, Gabriela. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina Fil: Taboga, Oscar Alberto. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina Fil: Del Vas, Mariana. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina 2023-09-12T13:57:27Z 2023-09-12T13:57:27Z 2011-10 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://hdl.handle.net/20.500.12123/15186 https://resjournals.onlinelibrary.wiley.com/doi/10.1111/j.1365-2583.2011.01100.x 1365-2583 https://doi.org/10.1111/j.1365-2583.2011.01100.x eng info:eu-repo/semantics/restrictedAccess http://creativecommons.org/licenses/by-nc-sa/4.0/ Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0) application/pdf The Royal Entomological Society Insect Molecular Biology 20 (5) : 675-685 (octubre 2011) |
| spellingShingle | Vectors Genomics Insects Viruses Vectores Genómica Insectos Virus Mal de Río Cuarto virus Virus del Mal de Río Cuarto Maroniche, Guillermo Andrés Mongelli, Vanesa Claudia Alfonso, Victoria Llauger, Gabriela Taboga, Oscar Alberto Del Vas, Mariana Development of a novel set of Gateway-compatible vectors for live imaging in insect cells |
| title | Development of a novel set of Gateway-compatible vectors for live imaging in insect cells |
| title_full | Development of a novel set of Gateway-compatible vectors for live imaging in insect cells |
| title_fullStr | Development of a novel set of Gateway-compatible vectors for live imaging in insect cells |
| title_full_unstemmed | Development of a novel set of Gateway-compatible vectors for live imaging in insect cells |
| title_short | Development of a novel set of Gateway-compatible vectors for live imaging in insect cells |
| title_sort | development of a novel set of gateway compatible vectors for live imaging in insect cells |
| topic | Vectors Genomics Insects Viruses Vectores Genómica Insectos Virus Mal de Río Cuarto virus Virus del Mal de Río Cuarto |
| url | http://hdl.handle.net/20.500.12123/15186 https://resjournals.onlinelibrary.wiley.com/doi/10.1111/j.1365-2583.2011.01100.x https://doi.org/10.1111/j.1365-2583.2011.01100.x |
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