TABLE 1.
Bacterial strains and plasmids
| Strain or plasmid | Relevant genotype | Reference or source |
|---|---|---|
| Strains | ||
| S. meliloti | ||
| Rm1021 | SU47 str-21, Smr | 30 |
| Rm5000 | SU47 rif-5, Rfr | 12 |
| Rm5439 | Rm1021 pckA1::TnV | 16 |
| SRmA185 | Rm1021 tpiA1::Tn5 | This work |
| SRmA327 | tpiA1, Φ(SRmA185)→Rm1021, Nmra | This work |
| SRmA355 | Rm5000 tpiB1::pKNOCK-Gm | This work |
| SRmA366 | tpiA1 tpiB1, Φ(SRmA185)→SRmA355, Nmr | This work |
| SRmA449 | Rm1021 ΔtpiA | This work |
| SRmA515 | ΔtpiA tpiB1, Φ(SRmA355)→SRmA449, Gmr | This work |
| SRmA584 | Rm1021 tpiB2::Tn5-B20 | This work |
| SRmA585 | ΔtpiA tpiB2, Φ(SRmA584)→SRmA449, Nmr | This work |
| E. coli | ||
| MM294A | pro-82 thi-1 hsdR17 supE44 | 14 |
| MT607 | MM294A recA56 | 14 |
| MT616 | MT607(pRK600) | 14 |
| S17-1 | recA derivative of MM294A with integrated RP4-2 (Tc::Mu::Km::Tn7) | 46 |
| DH5α | gyrA96, Nalr | Invitrogen |
| EcA100 | MT607 Rifr derivative | Lab collection |
| EcA101 | MT607 Tn5-B20 Kanr | 9 |
| Plasmids | ||
| pRK600 | pRK2013 npt::Tn9, Cmr | 14 |
| pRK602 | pRK600W Tn5, Cmr Nmr | 13 |
| pBluescriptII SK+ | Cloning vector, Apr | Stratagene |
| pKNOCK-Gm | Suicide vector, Gmr | 1 |
| pRK7813 | Broad-host-range vector, Tcr | 25 |
| pJQ200SK | Gene replacement suicide vector, Gmr | 42 |
| pPH1JI | IncP plasmid, Gmr Tcr | 5 |
| pNP150 | pKNOCK-Gm with 350-bp internal tpiB fragment | This work |
| pNP152 | pBluescriptII SK+ with 700-bp tpiA 5′ flanking region | This work |
| pNP154 | pBluescriptII SK+ with 660-bp tpiA 3′ flanking region | This work |
| pNP155 | pJQ200SK with tpiA flanking regions | This work |
| pNP163 | tpiB-complementing cosmid | This work |
| pNP166 | Constitutive tpiA | This work |
| pNP167 | Constitutive tpiB | This work |
a
Notation indicates that SRmA327 was constructed by ΦM12 transduction of SRmA185 into Rm1021 with selection for Nmr.