Figure 2.

Creating a functional tRNA_CUA^Pyl in yeast. (A) The consensus A- and B-box sequences and the A- and B-box sequences of MbtDNA_CUA. (B) The MbtDNA_CUA^Pyl expression constructs created and examined in this work. Constructs 6a−d were created using the 5′ and 3′ flanks from distinct tRNAs as described in the text. (C) Northern blots for MbtDNA_CUA expression from various constructs. (D) Phenotyping constructs for amber suppression in MaV203:pGADGAL4(2TAG) cells, where 3AT is 3-aminotriazole and 1 was used at 2 mM. Cells contained MbPylRS and the appropriate MbtDNA_CUA^Pyl expression construct.