Figure 2From: High-level HIV-1 Nef transient expression in Nicotiana benthamiana using the P19 gene silencing suppressor protein of Artichoke Mottled Crinckle VirusTime course expression analysis of Nef in vacuum agroinfiltrated N. benthamiana plants. Six weeks old Nicotiana benthamiana plants were vacuum-infiltrated with Agrobacterium. tumefaciens clones harbouring Nef, Nef/AMCV-P19, Nef/TBSV-P19 and Nef/PVX-P25. Leaf samples from three different plants were collected on 3, 5, 7, and 9 days post infiltration (d.p.i.). Leaf extracts were normalised for total soluble protein content (TSP) and analysed by direct ELISA using a mouse monoclonal antibody to HIV-1 Nef. For all samples a total of 50 μg of TSP were loaded in each ELISA plate well. C+: 100 ng of bacterially-derived Nef; C-: leaf extract of plants infiltrated with infiltration buffer. Values are the mean ± standard error of the mean (SEM) of triplicate samples.Back to article page