Table 2. Complementation of the mutation un-7.
| Transforming DNAa | Strain # (number of replicates) | Colonies at 37°C | HygR colonies at room temperature b |
| Protoplasts | |||
| pLORIST6xh H121G9 | 2176 (1) | 28 | 31 |
| pLORIST6xh H121G9 | 2175 (3) | 287 | ND |
| pMOcosX G14C6 | 2175 (3) | 38 | 20 |
| pMDFGSC651w | 2176 (2) | 45 | ND |
| pMDFGSC651c | 2176 (1) | 26 | ND |
| NCU00651 WT PCR | 2176 (1) | 20 | ND |
| NCU00651 9882 PCR | 2176 (4) | 41 | ND |
| No-DNA control c | 2175 (1) | 4 | 0 |
| No-DNA control | 2176 (2) | 12 | 0 |
| Electroporation | |||
| pLORIST6xh H121G9 | 2175 (2) | 54 | ND |
| pLORIST6xh H016E1 | 2175 (2) | 3 | >50 |
| pLORIST6xh H037A3 | 2175 (2) | 1 | ND |
| pLORIST6xh H011C11 | 2175 (2) | 1 | >100 |
| pLORIST6xh H110D8 | 2175 | 1 | ND |
| PCR NCU00651 | 2175 | 25 | ND |
| PCR NCU00652 | 2175 | 1 | ND |
| PCR NCU00653 | 2175 | 0 | ND |
| PCR NCU00654 | 2175 | 3 | ND |
| PCR NCU00655 | 2175 | 1 | ND |
Multiple transformations were carried out with each cosmid and the average result is presented for each strain tested.
a
In transformation with cosmid DNA, 3 ug of transforming DNA was used for each replicate. For transformation with PCR products, 0.8 to 1 ug of DNA was used for each experiment. DNA concentration was determined using a NanoDrop1000 UV/VIS Spectrophotometer.
b
Hygromycin phosphotransferase is carried on the pLORIST6xh and pMOcosX cosmids. This test was included as a control but was not possible with PCR products or genes cloned into pSC-A-amp/kan.
c
Colonies on no DNA control plates are the total number per 107–108 protoplasts.