PVX201 (which contains a DNA copy of the Potato Virus X genome with a duplicated sub-genomic promoter) and
pGR107 (PVX-based expression vector designed to combine the advantages of the A. tumefaciens-mediated transfection strategy and the infection power of the PVX virus) vectors were kindly provided from Prof. David Baulcombe (
The Sainsbury Laboratory, Norwich, UK) and PBL - Innovation in Life Sciences (
http://www.pbltechnology.com).
The PVX201 vector (kindly provided by David Baulcombe, The Sainsbury Laboratory, Norwich, UK) containing the PVX cDNA under the transcriptional control of the Cauliflower 35S promoter and the nopaline synthase terminator was ligated to the 2A double stranded oligolinker and then ligated to the 6His-tag oligolinker to produce the PVX6His2ACP vector. This vector contains a unique SalI site to be used for cloning coding sequences for the gene of interest that is fused in frame to the 3Õ terminus of the 6His-tag and to the 5Õ end of the 2A sequence.
