expression of a biotin acceptor peptide-containing protein with potential incorporation on the lentiviral envelope as a viral surface engineering platform

lentiviral vectors are among the promising viral based-vectors in gene therapy applications, but the efficiency of their targeting needs to be improved. (strept)avidin-biotin adaptor system is a novel approach to modify the lentiviral envelope for better targeting properties. herein, we describe utilization of this adaptor system by designing a candidate envelope protein-bearing biotin acceptor peptide (bap) and evaluation of its expression in 293t cells. to this end, a dna sequence containing flexible linkers, a 15-aminoacids bap and specific membrane regions of a viral protein was designed and synthesized in tandem. the synthesized gene was amplified with polymerase chain reaction to include bgl ii and sal i restriction sites and subcloned into the same sites of pdisplay vector in frame with ha-tag and myc epitope to construct the pdis-gs-bap. 293t cells were transfected with pdis-gs-bap and expression of resulting protein (dis-gs-bap) was evaluated by western blotting using anti-ha tag antibody. efficiency of transfection procedure was evaluated by pegfp-n1 vector and tracking for green fluorescent protein expression via fluorescence microscopy. restriction analysis and dna sequencing confirmed the precision of cloning steps. fluorescence microscopy indicated above 70% transfection efficiency and western blot analysis of pdis-gs-bap-transfected 293t cells showed a protein band of approximately 17 kda corresponding to the predicted size of dis-gs-bap protein. these promising results indicate the possibility of cell surface expression and further biotinylation of dis-gs-bap protein in ongoing studies.