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. 2011 May 17;67(Pt 6):560–567. doi: 10.1107/S0907444911014302

Figure 5.

Figure 5

DNA-replication activity of T-ag mutant proteins. (a) Transient DNA-replication activities of the indicated wt or mutant T-ag proteins in C33A cells. DNA-replication activities were measured by determining the ratio of firefly (Fluc-ori plasmid) to Renilla (Rluc control plasmid) luciferase activities as described in §2. Replication activities are reported as a percentage of the Fluc:Rluc ratio obtained with the largest amount of wt T-ag expression vector (12 ng). Cells transfected with vector only (No T-­ag) were used as a negative control. (b) Expression of T-ag proteins. Western blot analysis of total protein extracts prepared from transfected C33A cells expressing the wt or the indicated T-ag mutant proteins. Each protein was tested using a gradient of expression vector (0, 1.25, 2.5, 5.0, 10 and 12.5 ng). 100% replication was defined as the amount of replication observed using 12.5 ng wt T-ag.