Figure 2.

T7 transcription mediated by the s–Pa pairing. (A) Schemes of the experiments. (B) Gel electrophoresis of transcripts generated from templates containing one or two Pa or z bases with the natural NTPs (1 mM) and sTP (1 mM). Transcripts were labeled at their 5′-termini with [γ-³²P]GTP. The relative yields of each transcript were determined by comparison to the yields of native transcripts from templates consisting of the natural bases, and each yield was averaged from 3 to 4 data sets. (C) 2D-TLC analysis of the labeled ribonucleoside 3′-phosphates obtained from the nuclease digestion of the transcripts (17-mer). The transcripts were internally labeled with [α-³²P]UTP. The spots on the TLC were obtained from the 17-mer fragment transcribed from the template (N₁N₂N₃ = PaAC or CAC) in the presence of 1 mM sTP.