Figure 1
From: In vivo analysis of highly conserved Nef activities in HIV-1 replication and pathogenesis
A frame shift in nef ablates expression of Nef but does not affect viral replication. (A) Upper Panel, Left, Schematic representation of wild type LAI (WT LAI) is presented. Nucleotides 8390 to 9010 in NCBI accession number, K02013, represent the nef coding sequence. Lower Panel, Left, A schematic of frame shifted nef (LAINeffs) is presented. The insertion of GATC following nucleotide 8494 is indicated. PPT, polypurine tract. Upper Panel, Right, Flow chart describing the generation of the GATC insertion is presented. (B) The frame shift in nef eliminates Nef expression but does not alter the expression of Gag. nef (α-Nef) and gag (α-p24) encoded proteins were detected by Western blot analysis of 293T producer cell lysates. GAPDH (α-GAPDH) is a protein loading control. (C) A3.01 cells were infected with LAI or LAINeffs at a multiplicity of infection of 0.05 and viral production was followed for 20 days with ELISA for p24gag.