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. 2013 Jul 1;110(29):11803–11808. doi: 10.1073/pnas.1309584110

Fig. 4.

Fig. 4.

A single baculovirus vector encoding all of the genetic elements necessary to efficiently incorporate UAA into target proteins in mammalian cells. (A) pAcBac2 encodes a CAG promoter-driven reporter gene expression cassette, which also harbors a WPRE element at the 3′-UTR. Expression of aaRS is driven by a CMV promoter and lacks the WPRE element. (B) HEK293 cells infected with pAcBac2.tR4-OMeYRS/GFP* (MOI of 500) exhibits robust eGFP expression only in the presence of the UAA (1 mM). (C) FACS analysis of cells expressing eGFP in B. (D) Efficiency of GFP (Tyr39TAG) expression using pAcBac1.tR4-OMeYRS or pAcBac2.tR4-OMeYRS/GFP* are similar, when used at similar MOI (500). For expression using pAcBac.tR4-OMeYRS and-GFP*, the suppressor and the reporter viruses were used at the optimal 1:6 ratio or a 1:1 ratio (for a fixed total MOI of 500). Crude cell extracts were prepared 48 h after infection and GFP fluorescence was measured.