Fusion to Flaviviral Leader Peptide Targets HIV-1 Reverse Transcriptase for Secretion and Reduces Its Enzymatic Activity and Ability to Induce Oxidative Stress but Has No Major Effects on Its Immunogenic Performance in DNA-Immunized Mice

Figure 2

Expression of HIV-1 RT1.14 fused to the N-terminal signal peptide of NS1 protein of TBEV in HeLa cells resolved by SDS-PAGE. (a) Western blotting of the lysates of HeLa cells transfected with pVax1 (1), pVaxRT1.14opt-in (2), and pVaxRT1.14oil (3) plasmids; upper panel, membranes stained with anti-RT; lower panel, with anti-β-actin antibodies for signal normalization. (b) Western blotting of the culture fluids of cells transfected with pVax1 (1), pVaxRT1.14opt-in (2), and pVaxRT1.14oil (3). Blots were stained with the specific polyclonal anti-RT antibodies as described in the experimental section “Materials and Methods.” To control equal loading of the samples of cell lysates on the gel, membranes were for the second time stained with anti-β-actin antibodies. Positions of the molecular mass markers are given to the left.

(a)

(b)