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A simplified protocol for Agrobacterium-mediated transformation of cell suspension cultures of the model species Medicago truncatula A17
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Orientador(es)
Resumo(s)
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.
Descrição
Funding Information: Open access funding provided by FCT|FCCN (b-on). This work was supported by Fundação para a Ciência e Tecnologia (FCT, Portugal) through Research Units “Molecular, Structural and Cellular Microbiology” (MOSTMICRO, UIDB/ 04612/2020) and “Bioresources 4 Sustainability” (GREEN-IT, UIDB/04551/2020). Publisher Copyright: © 2023, The Author(s).
Palavras-chave
Agrobacterium Cultured cells Medicago Model legume Plant-based system RBD SARS-CoV-2 Stable transformation Horticulture