TABLE 1.
Competition efficiencies of circular and linearized ssDNA and dsDNAa
| Protein and competitor DNAc | Mean IC50 (nM)b ± SD
|
|
|---|---|---|
| ssDNA | dsDNA | |
| Pol/UL42 | ||
| Circular | 0.4 ± 0.006 | 0.07 ± 0.005 |
| Linear | 0.3 ± 0.05 | 0.02 ± 0.003 |
| Pol | ||
| Circular | 0.09 ± 0.006 | 2 ± 0.12 |
| Linear | 0.09 ± 0.02 | 0.5 ± 0.006 |
| UL42 | ||
| Circular | 0.5 ± 0 | 0.01 ± 0.0006 |
| Linear | 0.5 ± 0.01 | 0.01 ± 0.0006 |
a
A 15-fmol sample of a radiolabeled ds template (∼100 bp) was incubated with 0.5 pmol of Pol/UL42, Pol, or UL42 and various amounts of competitor DNA in 10-μl reaction mixtures in the presence of magnesium as detailed in Materials and Methods. The amount of radioactivity that bound to filters was used to deduce the IC50 of each competitor.
b
Data from three separate experiments are presented.
c
M13mp18 DNA was used as the competitor. The circular ssDNA and dsDNA used were positive-strand and RFI M13mp18, respectively. Linearized dsDNA was generated by restriction of RFI M13mp18 with EcoRI. Linearized ssDNA was obtained by heat denaturation of this restricted DNA.