We have developed a Potato virus X (PVX)-based vector system compatible with the GoldenBraid 2.0 (GB) cloning strategy to transiently express heterologous proteins or peptides in plants for biotechnological purposes. This vector system consists of three domestication vectors carrying three GB parts-the cauliflower mosaic virus (CaMV) 35S promoter with PVX upstream of the second subgenomic promoter of the PVX coat protein (PVX CP SGP), nopaline synthase (NOS) terminator with PVX downstream of the first PVX CP SGP and the gene of interest (GOI). The full-length PVX clone carrying the sequence encoding a green fluorescent protein (GFP) as GOI was incorporated into the binary GB vector in a one-step reaction of three GB parts using the four-nucleotide GB standard syntax. We investigated whether the obtained vector named GFP/pGBX enables systemic PVX infection and expression of GFP in Nicotiana benthamiana plants. We show that this GB-compatible vector system can be used for simple and efficient assembly of PVX-based expression constructs and that it meets the current need for interchange of standard biological parts used in different expression systems.

Department of Experimental Plant Biology Faculty of Science Charles University Prague 12843 Prague Czech Republic

Department of Plant Protection Czech University of Life Sciences 16500 Prague Czech Republic

Laboratory of Virology Centre for Plant Virus Research Institute of Experimental Botany of the Czech Academy of Sciences 16500 Prague Czech Republic

Plant Virus and Vector Interactions Centre for Plant Virus Research Crop Research Institute 16106 Prague Czech Republic

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