TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid/vector | Relevant feature(s) | Source or reference |
|---|---|---|
| Bradyrhizobium japonicum strains | ||
| USDA110 | Cmr (wild type) | 28 |
| 5051 | 110 (idhA::str), Cmr Strr | This study |
| 5051(pRS7) | 5051, Cmr Strr Tetr | This study |
| Escherichia coli strains | ||
| DH5α | supE44 Δ(lacZYA-argF)U169 deoR (Φ80lacZΔM15) gyrA98 endA1 hsdR17(rK− mK+) recA1 thi-1 relA1 | 19 |
| HB101 | thi-1 hsdR20(rB− mB+) supE44 lacY1 ara14 recA13 leuB6 proA2 xly5 mtl-1rpsL20 (Strr) | 4 |
| INVαF′ | F′ endA1 recA1 supE44 deoR Δ(lacZYA-argF)U169 λ−relA1 (Φ80lacZΔM15) hsdR17(rK− mK+) recA1 endA1 gyrA96 | Invitrogen |
| Plasmids/vectors | ||
| pTZ18U | Ampr, lacZ′, f1 ori, MCS, Mob−, cloning vector | 42 |
| pCR 2.1 | Ampr Kmr, lacZ′, f1 ori, MCS, Mob−, T cloning vector | Invitrogen |
| pGEM-T Easy | Ampr, lacZ′, f1 ori, MCS, Mob−, T cloning vector | Promega |
| pLO1 | Kmr, sacB, RP4 oriT, ColE1 ori | 30 |
| pHP45Ω | Ampr Strr in pP45Ω replicon | 42 |
| pRK2013 | Kmr, Mob+, Tra+, ColE1 ori, helper plasmid | 11 |
| pLAFR5 | Tcr, cosmid vector | 8 |
| pRS2 | Ampr, 2.01-kb PCR-amplified idhA region in pGEM-T Easy vector | This study |
| pRS3 | Ampr Strr, 2.01-kb PCR-amplified idhA region with a 270-bp XmaI deletion and a 2-kb XmaI Ω cassette insertion in pGEM-T Easy vector | This study |
| pRS4 | Ampr Strr, 3.73-kb PCR-amplified idhA::str insert with XbaI ends in pGEM-T Easy vector | This study |
| pRS5 | Kanr Strr, 3.73-kb PCR-amplified idhA::str insert in pLO1 vector | This study |
| pRS6a | Ampr Kanr, 2.01-kb PCR-amplified idhA region in pCR 2.1 vector | This study |
| pRS6b | Ampr Kanr, 2.01-kb PCR-amplified idhA region in pGEM-T Easy vector | This study |
| pRS7 | Tetr Kanr, 4.1-kb PCR-amplified idhA region and kan cassette with HindIII ends in pLAFR5 | This study |