Figure 5
Effect of TAE1 deletion on translation fidelity. (A) Deletion of TAE1 resulted in increased levels of β-gal activity from the lacZ reporters, pUKC817 and pUKC818, containing premature stop codons, UAA and UAG, respectively. The relative β-gal activity is determined by normalizing the activity of the mutant constructs (pUKC817 and pUKC818) to the control construct (pUKC815), and related to that obtained by the wild type strain (for example, the value for pUKC817/pUKC815 in tae1Δ cells is related to that of pUKC817/pUKC815 in the wild type cells). The average β-gal activity for the wild type strain transformed with pUKC815 is 19.05 (± 1.1) units. (B) Q-RT-PCR analysis indicates that alterations for the relative contents of LacZ mRNAs do not explain the difference in β-gal activities observed in (A). The Ct for the control and experimental samples were calculated from the threshold cycles. pUKC815 is the background construct without a premature stop codon and used as a control.