Figure 6
From: Expression, intracellular targeting and purification of HIV Nef variants in tobacco cells
Analysis and affinity purification of plant-produced cytosolic Nef. A: Samples of leaf protein homogenates from a p25 (line 120) and a p27 mut (line 116) transgenic line were resolved by 15% SDS-PAGE and subjected to immunoblot with anti-FLAG polyclonal antiserum. A dilution series of a recombinant, FLAG-tagged ScFv [28] was loaded as a quantitative reference. The asterisk indicates a non-specific, anti FLAG – immunoreactive peptide. Numbers at left indicate molecular weight markers in kDa. Co: control mock-transformed plant. B: Total leaf protein homogenate from transgenic plant line 116 expressing p27 mut was subjected to affinity chromatography with a cobalt resin. His-tagged Nef was eluted with imidazole. U: fraction not bound to the resin; W: wash fraction. Numbers at left indicate molecular weight markers in kDa.