Figure 4

Mutational analysis of the putative catalytic and zinc-binding residues of G1L utilizing a trans complementation assay. TREx 293 cells were transfected with 1 μg plasmid DNA bearing either wild type G1L or one of the single-site mutants. Four hours later the transfection solution was removed and replaced with infection media containing vvtetO:G1L at an MOI of 0.1 Cells were harvested at twenty-four hours post infection, subjected to a series of rapid freeze/thaws and titered via plaque assay on BSC₄₀ cells. Bars represent the percent rescue of each construct relative to what was achieved by transfection with the wild type G1L construct (G1L). Each transfection was carried out in the absence of TET (TET-).