Ferreira Ana Clara, Rebelo Bárbara A, Abranches Rita
Plant Cell Biology Laboratory, Instituto de Tecnologia Química e Biológica António Xavier, ITQB NOVA - Universidade Nova de Lisboa, Av. República, 2780-157 Oeiras, Portugal.
Plant Cell Tissue Organ Cult. 2023;153(3):669-675. doi: 10.1007/s11240-023-02495-6. Epub 2023 Mar 23.
This manuscript describes a unique protocol for the rapid transformation of A17 cell suspension cultures mediated by . 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.
本手稿描述了一种独特的方案，用于由……介导的A17细胞悬浮培养物的快速转化。在生长曲线的第7天收集苜蓿细胞，这对应于指数期的开始。然后将它们与农杆菌共培养3天，再铺在含有适当抗生素选择的培养皿上。使用严重急性呼吸综合征冠状病毒2（SARS-CoV-2）刺突蛋白的受体结合域作为模型来制定此方案。使用聚合酶链反应（PCR）评估转基因的存在，并通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）和蛋白质免疫印迹法（Western-blotting）评估产物的完整性。
