FIGURE 1.

Details of gene constructs. pcMT-full contained the rat MT-1 coding region and 3′UTR (Nury et al. 2005). pcMT-111 was made by PCR cloning (see Materials and Methods). Site-directed mutagenesis was used to delete successive 11–13-nucleotide regions of the MT-1 3′UTR to create pcMT-Δ1–13, -Δ14–26, -Δ26–36, -Δ36–46, -Δ46–56, -Δ56–66, -Δ66–76 (Nury et al. 2005), and -Δ76–86 (Nury et al. 2005), as well as nt 67–70 (first CACC motif) and 73–76 (second CACC motif) to produce pcMT-Δ(CACC₁) and pcMT-Δ(CACC₂), respectively. Site-directed mutagenesis was also used for substitutions of nt 21–25 (Nury et al. 2005), 26–30 (Nury et al. 2005), 67–70, and 74–75 of MT-1 3′UTR so as to create pcMT-S21–25, -S26–30, -S67–70, and -S74–75, respectively. Two double substitutions were made in which either both nt 27–30 and 67–70 or nt 26–31 and 66–71 were mutated. Details of all the sequence substitutions are shown in the figure.