TABLE 4.
Effect of glnL* alleles on the expression of the nitrogen regulated glnK promoter under nitrogen-limiting and nitrogen-replete conditions in a nac background
| Straind | Relevant genotypeb | β-Galactosidase activitya on:
|
|
|---|---|---|---|
| GGlnTrpc | GNGlnTrpc | ||
| YMC10φ | Wild type | 2,340 (1) | 0 (1) |
| Nφ | nac::Camr | 1,900 (1) | 0 (1) |
| BKgNφ | ΔglnB ΩGmrΔ glnK1 nac::Camr | 7,340 (1) | 5,830 (1) |
| Lφ | ΔglnL 2001 | 400 (1) | 60 (1) |
| L*(I12F)Nφ | glnL (I12F) nac::Camr | 4,800 | 850 |
| L*(L16R)Nφ | glnL (L16R) nac::Camr | 5,830 | 4,930 |
| L*(R116C)Nφ | glnL (R116C) nac::Camr | 4,750 | 1,020 |
| L*(A129T)Nφ | glnL (A129T) nac::Camr | 4,700 | 1,900 |
| L*(L154R)Nφ | glnL (L154R) nac::Camr | 5,410 | 4,500 |
| L*(G187R)Nφ | glnL (G187R) nac::Camr | 4,540 | 3,100 |
| YMC10φ | Wild type | 1,330 | 0 |
| Nφ | nac::Camr | 1,390 | 0 |
| BKgNφ | ΔglnB ΩGmrΔ glnK1 nac::Camr | 5,540 | 3,380 |
| L*(D225V)Nφ | glnL (D225V) nac::Camr | 5,180 | 3,480 |
| L*(S227N)Nφ | glnL (S227N) nac::Camr | 3,990 | 2,550 |
| L*(S227R)Nφ | glnL (S227R) nac::Camr | 5,170 | 3,780 |
| L*(S227I)Nφ | glnL (S227I) nac::Camr | 4,630 | 3,240 |
| L*(L228R)Nφ | glnL (L228R) nac::Camr | 5,130 | 4,050 |
| L*(Δ267-270)Nφ | glnL (Δ267-270) nac::Camr | 3,100 | 1,660 |
| L*(Y302N)Nφ | glnL (Y302N) nac::Camr | 4,200 | 2,900 |
| L*(P303L)Nφ | glnL (P303L) nac::Camr | 3,440 | 1,600 |
| L*(P303T)Nφ | glnL (P303T) nac::Camr | 4,530 | 3,530 |
β-Galactosidase activities are given in Miller units and are the average for duplicate cultures except where indicated by the number of cultures in parentheses. Results from duplicate cultures differed by <10%. Cultures were grown overnight in the indicated medium, diluted to an optical density at 600 nm of 0.02, and grown at 30°C to an optical density of 0.5.
In addition to the genotype shown, all strains contain trpDC700::putPA1303 [Kanr glnKp-lac], which is a fusion of the lacZYA operon to the nitrogen regulated glnK promoter.
Media used were G GlnTrp (glucose-glutamine-tryptophan) and GN Gln Trp (glucose-ammonia-glutamine-tryptophan) and contained kanamycin. Glucose was present at 0.4% (wt/vol), ammonia and glutamine were present at 0.2% (wt/vol), and tryptophan was present at 0.04 mg/ml.
The break in the table indicates experiments performed on separate days.