Table 1.
Nucleotide-composition analysis of T7 transcripts
| Entry | Template N1N2N3 | [α-32P] NTP | sTP or s′TP (mM) | Composition of nucleotides incorporated as 5′ neighbor of U or A or Ga | ||||
|---|---|---|---|---|---|---|---|---|
| Ap | Gp | Cp | Up | s′p | ||||
| 1 | PaAC | UTP | sTP (1) | 0.01b [0]c (0.01)d | n.d.e [0] (<0.01) | 1.99 [2] (0.05) | 1.02 [1] (0.03) | 0.97 [1] (0.03) |
| 2 | zAC | UTP | sTP (1) | 0.01 [0] (<0.01) | 0.01 [0] (0.01) | 1.99 [2] (0.02) | 1.02 [1] (0.05) | 0.97 [1] (0.05) |
| 3 | CAC | UTP | – (0) | 0.01 [0] (<0.01) | 0.97 [1] (0.03) | 2.02 [2] (0.02) | 1.01 [1] (0.03) | n.d. [0] (–) |
| 4 | CAC | UTP | sTP (1) | 0.01 [0] (<0.01) | 0.98 [1] (0.02) | 2.03 [2] (0.03) | 0.99 [1] (0.04) | n.d. [0] (–) |
| 5 | TAC | UTP | – (0) | 1.00 [1] (0.03) | 0.01 [0] (<0.01) | 2.02 [2] (0.01) | 0.97 [1] (0.04) | n.d. [0] (–) |
| 6 | TAC | UTP | sTP (1) | 1.00 [1] (0.03) | 0.01 [0] (<0.01) | 2.01 [2] (0.01) | 0.97 [1] (0.03) | 0.01 [0] (<0.01) |
| 7 | PaAPa | UTP | sTP (1) | 0.01 [0] (0.01) | 0.01 [0] (<0.01) | 2.03 [2] (0.03) | 1.00 [1] (0.02) | 0.95 [1] (0.03) |
| 8 | PaAPa | UTP | sTP (3) | 0.01 [0] (0.01) | 0.01 [0] (0.01) | 2.01 [2] (0.01) | 0.99 [1] (0.01) | 0.98 [1] (0.01) |
| 9 | PaAPa | ATP | sTP (1) | 0.05 [0] (<0.01) | 1.06 [1] (0.07) | n.d. [0] (–) | 0.01 [0] (<0.01) | 0.87 [1] (0.07) |
| 10 | PaAPa | ATP | sTP (3) | 0.04 [0] (0.01) | 1.02 [1] (0.06) | n.d. [0] (–) | 0.01 [0] (<0.01) | 0.93 [1] (0.07) |
| 11 | CAC | ATP | sTP (1) | 0.03 [0] (<0.01) | 1.95 [2] (0.01) | n.d. [0] (–) | 0.02 [0] (0.01) | n.d. [0] (–) |
| 12 | CAC | ATP | sTP (3) | 0.03 [0] (<0.01) | 1.94 [2] (0.01) | n.d. [0] (–) | 0.02 [0] (0.01) | n.d. [0] (–) |
| 13 | PaPaC | GTP | sTP (1) | 1.99 [2] (0.05) | 1.15 [1] (0.05) | 0.02 [0] (0.02) | n.d. [0] (–) | 0.84 [1] (0.03) |
| 14 | PaPaC | GTP | sTP (3) | 1.94 [2] (0.03) | 1.13 [1] (0.07) | 0.02 [0] (0.01) | 0.01 [0] (0.02) | 0.89 [1] (0.10) |
| 15 | CAC | GTP | sTP (1) | 1.98 [2] (0.06) | 1.01 [1] (0.03) | 0.01 [1] (0.01) | 1.98 [2] (0.08) | 0.01 [1] (0.01) |
| 16 | CAC | GTP | sTP (3) | 1.99 [2] (0.04) | 0.99 [1] (0.03) | 0.01 [0] (0.01) | 1.99 [2] (0.08) | 0.02 [1] (0.01) |
| 17 | PaAC | UTP | s′TP (1) | 0.02 [0] (<0.01) | 0.01 [0] (0.01) | 2.01 [2] (0.02) | 1.01 [1] (0.01) | 0.95 [1] (0.01) |
| 18 | zAC | UTP | s′TP (1) | 0.04 [0] (<0.01) | 0.08 [0] (0.01) | 2.01 [2] (0.01) | 1.01 [1] (0.01) | 0.85 [1] (0.01) |
| 19 | CAC | UTP | s′TP (1) | 0.01 [0] (<0.01) | 0.98 [1] (0.01) | 2.01 [2] (0.02) | 1.00 [1] (0.02) | n.d. [0] (–) |
aComposition of nucleotides incorporated as 5′ neighbor of U (Entries 1–8 and 17–19), A (Entries 9–12) or G (Entries 13–16), as shown in Figure 2.
bThe values were determined using the following formula: (radioactivity of each nucleotide)/[total radioactivity of all nucleotides (3′-monophosphates)] × (total number of nucleotides at 5′ neighbor of [α-32P]NTP).
cThe theoretical number of each nucleotide is shown in brackets.
dSDs are shown in parentheses.
eNot detected.