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. 2001 Feb 6;98(4):1537–1542. doi: 10.1073/pnas.031561698

Figure 1.

Figure 1

(A) A brief description of mutants made in this paper. Rat PAH coding sequence was cloned in to an expression vector pET26b(+) (Novagen), which contains a T7 promoter sequence (T7). The broken bold line represents part of the resulting plasmid pET26-rPAH, corresponding to the 5′ coding sequence. The numbers under the line indicate the position of nucleotides; the initiation codon starts with number 1. (Bottom) The first 40 amino acids from rat (RAT) and human PAH (HUMAN). Shaded areas represent identical sequences in rat and human PAH. The primers (arrows) and restriction sites used for mutagenesis are shown at relative positions. (Middle) Amino acids at site 29 of rat PAH and their corresponding enzyme names. wt, wild type. (B) Primer usage for the first round of PCR as described in Materials and Methods. a, primers in combination with primer o265; b, primers in combination with primer N-rev; c, the template was human PAH sequence.