TABLE 1.
Wild-type EBNA-1 and NΔ330-641 support replication of plasmids oriP-BamHI C-Luc and FR-BamHI C-Luc but not DS-BamHI C-Luc in 143B cells at 96 h postelectroporation
| Effector | Relative efficiency of replicationa
|
||
|---|---|---|---|
| oriP-BamHI C-Luc | FR-BamHI C-Luc | DS-BamHI C-Luc | |
| Vector | <2.1 | <2.1 | <2.1 |
| EBNA-1 | 100 | 43 ± 3.4 | <2.1 |
| NΔ330-641 | 18 ± 0.94 | 2.3 ± 0.19 | <2.1 |
10 μg of each DNA (effector, reporter, and oriP-minus) was introduced into 107 143B cells by electroporation. Low-molecular-weight DNA was isolated after 94 to 98 h, and the relative concentration of DpnI-resistant plasmids was determined by quantitative competitive PCR and was corrected for transfection efficiency of 143B cells as described in Materials and Methods. The relative efficiency of replication by each derivative of EBNA-1 is expressed as a percentage of DpnI-resistant DNA relative to that of oriP-BamHI C-Luc replicated by wild-type EBNA-1, which is set to 100% (100% = 16 ± 14 copies of DpnI-resistant DNA per transfected cell). Data represent an average of two experiments ± standard deviation. The level of DpnI-resistant oriP-minus DNA detected was less than the lowest amount competitor used in all cases: <8.8% when oriP-BamHI C-Luc, oriP-minus, and wild-type EBNA-1 were cotransfected and <2.1% for all other reporter-effector combinations. Data represented as “<” indicate that the level of DpnI-resistant DNA was less than the smallest amount used for the competitor DNA and is therefore reported as less than an average of the smallest amount per transfected cell that could be determined by interpolation from the competitor DNA curve.