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

Abstract

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.

Performance of plant-produced RBDs as SARS-CoV-2 diagnostic reagents: a tale of two plant platforms

“…Plant-based recombinant COVID-19 works have mostly been based on transient expression in Nicotiana benthamiana (see Table 1 and references therein). Only few studies involved the stable transformation of plant cell suspensions ( Daniell et al., 2022 ; Rebelo et al., 2022 ; Ferreira et al., 2023 ; Shen et al., 2023 ). Among the proteins reported, there are structural proteins such as the spike (S) protein or S subunits (S1 and S2), the receptor-binding domain (RBD), the nucleocapsid (NC), envelope (E), and membrane (M) viral proteins, and others such as angiotensin-converting enzyme 2 (ACE2) receptor and antibodies against domains of the spike protein (RBD and S1) and the nucleocapsid protein ( Table 1 ).…”