Table 1.
ϕ29 TP | TP-dAMP formationa |
TP-DNA replicationa | Transitiona,c | TP-DNA amplification |
||
---|---|---|---|---|---|---|
TP-DNAb | No template | (%)a Amplification factord | ||||
wild-type | 100 | 100 | 100 | 1 | 100 | 50 ± 7 |
R158A | 4 ± 1.5 | n.d. | 17 ± 3 | n.d. | 1 ± 0.2 | 1 ± 0.1 |
R169A | 23 ± 6 | n.d. | 75 ± 13 | 1.7 ± 0.4 | 3 ± 0.3 | 3 ± 0.2 |
E191A | 94 ± 11 | 30 ± 8 | 106 ± 17 | 2.2 ± 0.3 | 69 ± 2 | 35 ± 6 |
D198A | 112 ± 1 | 102 ± 3 | 66 ± 8 | 0.3 ± 0.2 | 121 ± 10 | 61 ± 13 |
Y250A | 74 ± 24 | 10 ± 2 | 100 ± 20 | 3.3 ± 0.4 | 130 ± 4 | 66 ± 10 |
E252A | 70 ± 28 | n.d. | 103 ± 25 | 3.2 ± 0.8 | 52 ± 7 | 27 ± 7 |
Q253A | 23 ± 5 | n.d. | 55 ± 12 | 4.7 ± 0.7 | 20 ± 6 | 10 ± 4 |
R256A | 20 ± 8 | n.d. | 76 ± 19 | 2.6 ± 1.2 | 16 ± 5 | 10 ± 1 |
Data represent the mean value and the standard deviation obtained from at least three independent experiments.
aNumbers indicate the activity of mutant TPs with respect to the wild-type TP.
bTP-dAMP formation using ϕ29 TP-DNA as template of the reaction.
cTransition efficiency was calculated as the ratio of the truncated products TP-(dNMP)6-14/TP-(dNMP)1–2 considering such quotient 1 in the case of wild-type TP. See details in the main text.
dThe amplification factor was calculated as the ratio between the amount of DNA at the end of the reaction (input plus synthesized DNA) and the amount of input DNA.
n.d. Not detected.