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

Fig. 2.

Fig. 2.

Baculovirus vectors encoding orthogonal tRNA/aaRS pairs. (A) Two different suppressor tRNAs (both for the tyrosine and the pyrrolysine system) were expressed from two different promoters (U6 and H1). (B) Structure of pAcBac1.tR4, which encodes the two-copy tRNA cassette in two different orientations. Either the aaRS or the reporter gene eGFP (Tyr39TAG) was inserted into the multiple cloning site (MCS), downstream of the CMV promoter to generate the suppressor or the reporter virus, respectively. (C) Structures of pAcF and eBK. (D) Expression of eGFP (Tyr39TAG) in HEK293 cells by amber suppression using pAcBac1.tR4-OMeYRS (Upper) or pAcBac1.tR4-MbPylRS (Lower), in the presence or absence of 1 mM pAcF or eBK, respectively. (E) FACS analysis of cells expressing eGFP (Tyr39TAG) as described in D.