A simplified protocol for Agrobacterium-mediated transformation of cell suspension cultures of the model species Medicago truncatula A17

This manuscript describes a unique protocol for the rapid transformation of Medicago truncatula A17 cell suspension cultures mediated by Agrobacterium tumefaciens. Medicago cells were collected on day 7 of the growth curve, which corresponded to the beginning of the exponential phase. They were then...

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Detalles Bibliográficos
Autores principales: Ferreira, Ana Clara, Rebelo, Bárbara A., Abranches, Rita
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Netherlands 2023
Materias:
Protocol Paper
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10034231/
https://www.ncbi.nlm.nih.gov/pubmed/37197004
http://dx.doi.org/10.1007/s11240-023-02495-6
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author Ferreira, Ana Clara
Rebelo, Bárbara A.
Abranches, Rita
author_facet Ferreira, Ana Clara
Rebelo, Bárbara A.
Abranches, Rita
author_sort Ferreira, Ana Clara
collection PubMed
description This manuscript describes a unique protocol for the rapid transformation of Medicago truncatula A17 cell suspension cultures mediated by Agrobacterium tumefaciens. Medicago cells were collected on day 7 of the growth curve, which corresponded to the beginning of the exponential phase. They were then co-cultured with Agrobacterium for 3 days before being spread onto a petri dish with appropriate antibiotic selection. The Receptor Binding Domain of the Spike protein of SARS-CoV-2 was used as a model to develop this protocol. The presence of the transgene was assessed using PCR, and the integrity of the product was evaluated by SDS-PAGE and Western-blotting.
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spelling pubmed-100342312023-03-23 A simplified protocol for Agrobacterium-mediated transformation of cell suspension cultures of the model species Medicago truncatula A17 Ferreira, Ana Clara Rebelo, Bárbara A. Abranches, Rita Plant Cell Tissue Organ Cult Protocol Paper This manuscript describes a unique protocol for the rapid transformation of Medicago truncatula A17 cell suspension cultures mediated by Agrobacterium tumefaciens. Medicago cells were collected on day 7 of the growth curve, which corresponded to the beginning of the exponential phase. They were then co-cultured with Agrobacterium for 3 days before being spread onto a petri dish with appropriate antibiotic selection. The Receptor Binding Domain of the Spike protein of SARS-CoV-2 was used as a model to develop this protocol. The presence of the transgene was assessed using PCR, and the integrity of the product was evaluated by SDS-PAGE and Western-blotting. Springer Netherlands 2023-03-23 2023 /pmc/articles/PMC10034231/ /pubmed/37197004 http://dx.doi.org/10.1007/s11240-023-02495-6 Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Protocol Paper
Ferreira, Ana Clara
Rebelo, Bárbara A.
Abranches, Rita
A simplified protocol for Agrobacterium-mediated transformation of cell suspension cultures of the model species Medicago truncatula A17
title A simplified protocol for Agrobacterium-mediated transformation of cell suspension cultures of the model species Medicago truncatula A17
title_full A simplified protocol for Agrobacterium-mediated transformation of cell suspension cultures of the model species Medicago truncatula A17
title_fullStr A simplified protocol for Agrobacterium-mediated transformation of cell suspension cultures of the model species Medicago truncatula A17
title_full_unstemmed A simplified protocol for Agrobacterium-mediated transformation of cell suspension cultures of the model species Medicago truncatula A17
title_short A simplified protocol for Agrobacterium-mediated transformation of cell suspension cultures of the model species Medicago truncatula A17
title_sort simplified protocol for agrobacterium-mediated transformation of cell suspension cultures of the model species medicago truncatula a17
topic Protocol Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10034231/
https://www.ncbi.nlm.nih.gov/pubmed/37197004
http://dx.doi.org/10.1007/s11240-023-02495-6
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