TABLE 3.
Hybridization profiles from Southern analyses of wild-type and mutant strains
| Mutant | Restriction endonuclease(s) | Probea | Hybridizing fragment (kb)
|
|
|---|---|---|---|---|
| Parentb | Mutant | |||
| ceaS2-Fs | EcoRI/NruI | ceaS2 | 3.5 | 2.0 |
| apr | 3.5 | None | ||
| ΔceaS1::apr | NcoI | ceaS1 | 1.2c | Nonec |
| apr | None | 3.4 | ||
| ΔceaS1::apr/ceaS2-Fs | NcoI | ceaS1 | 1.2c | Nonec |
| apr | None | 3.4 | ||
| bls1::tsr | NcoI | bls1 | 1.9c | 3.0c |
| tsr | None | 3.0 | ||
| bls1::tsr/bls2::apr | NcoI | bls1 | 1.9c | 3.0c |
| tsr | None | 3.0 | ||
| oat1::neo | BglII | oat1 | 4.3 | 2.4 and 2.9 |
| neo | None | 2.4 and 2.9 | ||
| oat1::neo/oat2::apr | BglII | oat2 | 6.5 | 7.95 |
| neo | None | 7.95 | ||
Gene-specific probes are described in Materials and Methods. An approximately 1.5-kb fragment from pUC120Apr was used as the apramycin-specific probe. An approximately 1-kb EcoRI-HindIII fragment from pTSR#8 was used as the thiostrepton-specific probe. An approximately 1-kb BamHI-HindIII fragment from pFDNeo-S was used as the neomycin-specific probe.
ceaS2::apr was the parental strain for the ceaS2-Fs mutant, ceaS2-Fs was the parental strain for the ΔceaS1::apr/ceaS2-Fs double mutant, bls2::apr was the parental strain for the bls1::tsr/bls2::apr double mutant, and oat1::apr was the parental strain for the oat1::neo/oat2::apr double mutant. For all other mutants, the parental strain was the wild type.
Faint cross-hybridizing bands were observed which can be attributed to the presence of the respective paralogues.