Figure 2

Characterization of recombinant viruses with PBS complementary to tRNA^Met(e) and tRNA^Met(i). Panel A. Production of infectious virus following transfection of proviral plasmids. The designated proviral plasmids were transfected into 293T cells and the supernatant assayed for production of infectious virus using the JC53-BL assay. Culture volumes for each virus were the same. Error bars ± standard deviation. Panel B. p24 antigen production from transfected cells. Cells were transfected with different amounts of HXB2-WT, HXB2-Met(e) or HXB2-Met(i) and the p24 antigen in the culture supernatant was determined by solid phase ELISA. The amounts for each transfection was as follows: Lane 1 : 1 μg, Lane 2 : 2 μg, Lane 3 : 3 μg, Lane 4 : 4 μg, Lane 5 : 8 μg of proviral plasmid DNA. Panel C. Analysis of virus produced from transfected cells. Virus from transfected cells was pelleted by ultracentrifugation and subjected to SDS PAGE and Western blot using antibody specific for HIV-1 Gag. The order of the samples are as follows: Lane 1 – HXB2-Met(e), Lane 2 : HXB2-Met(i), Lane 3 : HXB2-WT. The positions of a viral gag gene products CA p24, p41 and pr55^Gag are noted.