Table 3.
Effect of subculture conditions on pyrF elimination and generation of false positives
| Subculture | Medium | Frequency of occurrence (%)a |
||
|---|---|---|---|---|
| 5-FOA resistantb | False positivec | Revertantd | ||
| Procedure A | ||||
| 1 | LB | 0.01 ± 0.00 | ||
| 2 | LB | 0.45 ± 0.42 | ||
| 3 | LB | 0.29 ± 0.28 | 63 ± 5 | |
| 4 | +5-FOAe | 59 ± 53 | 7.8 ± 6.0 | 61 ± 17 |
| Procedure B | ||||
| 1 | MMf | 4.5 ± 0.9 | ||
| 2 | MM | 5.5 ± 3.9 | ||
| 3 | MM | 5.5 ± 2.9 | 86 ± 9 | |
| 4 | +5-FOA | 9.4 ± 5.6 | 8.1 ± 13 | 68 ± 12 |
The values shown are means ± SD from four independent experiments using transconjugants that had integrated pGAM47-bgaB.
The values shown are the number of colonies growing from subculture aliquots on MM plates containing 10 mg/liter uracil and 5-FOA compared with the number on LB plates (i.e., 5-FOA-resistant cells per total cells).
The values shown are the number of clones growing on MM plates without uracil per 48 5-FOA-resistant clones (i.e., uracil-prototrophic cells per 5-FOA-resistant cells).
The values shown are the number of clones exhibiting white colonies on LB plates with 200 mg/liter X-Gal per 32 uracil-auxotrophic clones generated (i.e., MK72 revertant cells per total cells from which pyrF had been eliminated).
MM containing 1 mg/liter uracil and 5-FOA.
MM containing 10 mg/liter uracil.