Figure 1.

In vitro transcription of a U6 gene containing a variant TATA-box. In vitro transcription systems based on 10 µl phosphocellulose fraction B and 20 µl phosphocellulose fraction C (lanes 1–21) containing either pUC18 (lanes 1–7), pUhU6_0.35 (lanes 8–14) or pUhU6_{0.35 –29<<} (lanes 15–21) as template, were supplemented either with increasing amounts (5–20 ng) of recombinant human TBPwt (lanes 2–4, 9–11 and 16–18) or recombinant human TBP-DR2 (lanes 5–7, 12–14 and 19–21), respectively. Lanes 1, 8 and 15 contained no recombinant TBP. Lanes 2, 5, 9, 12, 16 and 19 were supplemented with 5 ng, lanes 3, 6, 10, 13, 17 and 20 with 10 ng, and lanes 4, 7, 11, 14, 18 and 21 with 20 ng recombinant TBPwt or TBP-DR2 respectively. In vitro transcription was carried out using 1 µg plasmid DNA as template. The position of the U6 transcripts is indicated and the position of the TBP-DR2 dependent transcript is marked with an asterisk.