Institute of Experimental Botany of the Czech Academy of Sciences, Prague, Czech Republic.
Biopharming Research Unit, Department of Molecular and Cell Biology, University of Cape Town, Cape Town, South Africa.
Plant Cell Rep. 2020 Sep;39(9):1115-1127. doi: 10.1007/s00299-020-02544-w. Epub 2020 Apr 24.
This is the first evidence that replicating vectors can be successfully used for transient protein expression in BY-2 plant cell packs. Transient recombinant protein expression in plants and recently also plant cell cultures are of increasing interest due to the speed, safety and scalability of the process. Currently, studies are focussing on the design of plant virus-derived vectors to achieve higher amounts of transiently expressed proteins in these systems. Here we designed and tested replicating single and multi-cassette vectors that combine elements for enhanced replication and hypertranslation, and assessed their ability to express and particularly co-express proteins by Agrobacterium-mediated transient expression in tobacco BY-2 plant cell packs. Substantial yields of green and red fluorescent proteins of up to ~ 700 ng/g fresh mass were detected in the plant cells along with position-dependent expression. This is the first evidence of the ability of replicating vectors to transiently express proteins in BY-2 plant cell packs.
这是首例证据表明，复制载体可成功用于 BY-2 植物细胞团的瞬时蛋白表达。瞬时重组蛋白在植物中的表达，以及最近在植物细胞培养物中的表达，由于该过程的速度、安全性和可扩展性而受到越来越多的关注。目前，研究的重点是设计植物病毒衍生载体，以在这些系统中实现更高水平的瞬时表达蛋白。在这里，我们设计和测试了复制的单盒和多盒载体，这些载体结合了增强复制和超翻译的元件，并通过农杆菌介导的瞬时表达在烟草 BY-2 植物细胞团中评估了它们表达和特别共表达蛋白的能力。在植物细胞中检测到高达约 700ng/g 鲜重的绿色和红色荧光蛋白的大量产量，以及位置依赖性表达。这是首例证据表明复制载体能够在 BY-2 植物细胞团中瞬时表达蛋白。
