Figure 2.

C-Ala is important for the aminoacylation activity of CeAlaRS_c. (A) Aminoacylation assay. The aminoacylation activities of CeAlaRS_c and CeAlaRS_c(ΔC-Ala) were determined using various tRNAs as substrates. (B) Complementation assay. The rescue activities of CeAlaRS_c and its C-terminal deletion construct were determined by transforming the test plasmid into a yeast ALA1 KO strain and plating the resultant transformants on 5-FOA. The symbols ‘+’ and ‘−’ denote positive and negative complementation, respectively. (C) Western blotting. The protein expression levels of these constructs were determined by western blotting using an anti-His₆-tagged antibody as the probe. Constructs used in (B) and (C) are numbered for clarity.