Table I.
Recovery of phleomycin-resistant transformants
| Construct |
nit1-305cw15
|
CC-3396
|
|||
|---|---|---|---|---|---|
|
KpnI-digested
DNA
|
KpnI-digested
DNA
|
Supercoiled DNA
|
|||
| 2 μg/mL Phl | 5 μg/mL Phl | 2 μg/mL Phl | 5 μg/mL Phl | 5 μg/mL Phl | |
| pSP109 | 13 | 13 | 15 | 7 | 4 |
| pγ-ble | 7 | 4 | 8 | 1 | 0 |
| pγ-ble Δ7-26 | 35 | 30 | 30 | 4 | 5 |
| pγ-ble Δ7-29 | 15 | 8 | 4 | 1 | 6 |
| pγ-ble Δ27-29 | 17 | 10 | 17 | 24 | 4 |
| −DNA | 0 | 0 | 0 | 0 | 0 |
Glass bead transformation was carried out with 2 μg of the indicated DNA and 4 × 107 cells (Kindle, 1990). Cells grew in acetate-containing liquid medium for 18 h following transformation, and cultures were split in half and plated on high-salt acetate (CC-3396) or SGII-NH4 (nit1-305cw15) plates containing 2 or 5 μg/mL phleomycin (Phl).