TABLE 1.
Bacterial strains, cosmids, and plasmids used in this study
| Strain, cosmid, or plasmid | Description | Source or reference |
|---|---|---|
| Strains | ||
| 3841 | Rhizobium leguminosarum bv. viciae Smr derivative of strain 300 | 24 |
| A34 | Rhizobium leguminosarum bv. viciae (formerly known as 8401 pR11JI) | 10 |
| VF39SM | Rhizobium leguminosarum bv. viciae Smr | 38 |
| RU1357 | A34 braE::TnphoA ΔaapJQM::ΩSp | 20 |
| RU1180 | 3841 mctP::TnlacZ | This study |
| RU1580 | pRU895 homogenote in 3841; mctS::GeneJumper Kmr transposon | This study |
| RU1581 | pRU897 homogenote in 3841; mctP::GeneJumper Kmr transposon | This study |
| RU1582 | pRU898 homogenote in 3841; mctR::GeneJumper Kmr transposon | This study |
| RU1583 | pRU899 homogenote in 3841; mctR::GeneJumper Kmr transposon | This study |
| RU1584 | pRU900 homogenote in 3841; mctS::GeneJumper Kmr transposon | This study |
| RU1585 | pRU901 homogenote in 3841; mctR::GeneJumper Kmr transposon | This study |
| RU1586 | pRU902 homogenote in 3841; mctS::GeneJumper Kmr transposon | This study |
| RU1587 | pRU903 homogenote in 3841; mctS::GeneJumper Kmr transposon | This study |
| RU1601 | 3841 pRU923 | This study |
| RU1602 | RU1580 pRU923 | This study |
| RU1603 | RU1581 pRU923 | This study |
| RU1604 | RU1582 pRU923 | This study |
| RU1632 | RU1357 pRU976 | This study |
| RU1633 | RU1357 pRU986 | This study |
| Cosmid | ||
| pRU3147 | pLAFR1 cosmid containing mct region from strain 3841 | This study |
| Plasmids | ||
| pCR2.1 TOPO | Cloning vector for PCR products; Kmr, Ampr | Invitrogen Life Technologies |
| pCR-Blunt-TOPO | Cloning vector for PCR products; Kmr | Invitrogen Life Technologies |
| pDONR201 | PCR cloning vector for Gateway Technology | Invitrogen Life Technologies |
| pJQ200SK | pACYC derivative, P15A origin of replication; Gmr | 39 |
| pLAFR1 | Wide-host-range mobilizable P-group cloning vector; Tcr | 11 |
| pPHJI1 | P-group chaser plasmid | 16 |
| pRK415-1 | Broad-host-range P-group cloning vector; Tcr | 27 |
| pJP2 | pTR102 GUS (mini-RK2 derivative) with artificial MCS; Ampr Tcr | 37 |
| pMN402 | GFP+ expression vector; hygromycin resistance | 47 |
| pOT2 | Promoter probe vector with promoterless gfpUV; Gmr | 2 |
| pOT2GFP+ | Promoter probe vector with promoterless gfp+; Gmr | This study |
| pOT3GFPuv | pOT2 with Gateway vector conversion cassette (reading frame A; Invitrogen Life Technologies) cloned into SmaI site | This study |
| pOT3GFP+ | pOT2GFP+ with Gateway vector conversion cassette (reading frame A; Invitrogen Life Technologies) cloned into SmaI site | This study |
| pRU738 | 2-kb PCR product (primers P237 and P238; mctP) from pRU3147 cloned into pCR2.1-TOPO | This study |
| pRU739 | 2-kb EcoRI fragment from pRU738 in pRK415-1 with mctP under the control of the lac promoter | This study |
| pRU747 | 2-kb EcoRI fragment from pRU738 in pRK415-1 with mctP not under the control of the lac promoter | This study |
| pRU785 | 3.6-kb SalI fragment from pRU3147 in pRK415-1 with mctRP not under the control of the lac promoter | This study |
| pRU788 | 3.6-kb SalI fragment from pRU3147 in pRK415-1 with mctRP under the control of the lac promoter | This study |
| pRU887 | 4.8-kb PCR product (primers P238 and P285; mctSRP) from pRU3147 cloned into pCR2.1-TOPO | This study |
| pRU890 | pJQ200SK containing a 4.4-kb BamHI fragment from pRU887 | This study |
| pRU895 | pRU890 mctS::GeneJumper Kmr transposon | This study |
| pRU897 | pRU890 mctP::GeneJumper Kmr transposon | This study |
| pRU898 | pRU890 mctR::GeneJumper Kmr transposon | This study |
| pRU899 | pRU890 mctR::GeneJumper Kmr transposon | This study |
| pRU900 | pRU890 mctS::GeneJumper Kmr transposon | This study |
| pRU901 | pRU890 mctR::GeneJumper Kmr transposon | This study |
| pRU902 | pRU890 mctS::GeneJumper Kmr transposon | This study |
| pRU903 | pRU890 mctS::GeneJumper Kmr transposon | This study |
| pRU917 | PCR product (primers P316 and P317) containing the putative mctP promoter region from pRU3147 in pDONR201 | This study |
| pRU922 | mctP promoter region from pRU917 in pOT3GFPuv | This study |
| pRU923 | mctP promoter region from pRU917 in pOT3GFP+ | This study |
| pRU935 | 1.8-kb PCR product (primers P345 and P343; mctS) from pRU3147 cloned into pCR-Blunt II-TOPO | This study |
| pRU936 | 2.7-kb PCR product (primers P342 and P343; mctSR) from pRU3147 cloned into pCR-Blunt II-TOPO | This study |
| Strain, cosmid, or plasmid | Description | Source or reference |
| pRU941 | 2.3-kb PCR product (primers P340 and P341; mctP) from pRU3147 cloned into pCR2.1-TOPO | This study |
| pRU942 | 1.5-kb PCR product (primers P342 and P344; mctR) from pRU3147 cloned into pCR2.1-TOPO | This study |
| pRU947 | 2.3-kb HindIII fragment from pRU941 in pRK415-1 with mctP under the control of the lac promoter | This study |
| pRU948 | 2.3-kb HindIII fragment from pRU941 in pRK415-1 with mctP not under the control of the lac promoter | This study |
| pRU950 | 1.8-kb EcoRI fragment from pRU935 in pRK415-1 with mctS under the control of the lac promoter | This study |
| pRU951 | 1.5-kb EcoRI fragment from pRU942 in pRK415-1 with mctR under the control of the lac promoter | This study |
| pRU952 | 2.7-kb EcoRI fragment from pRU936 in pRK415-1 with mctSR under the control of the lac promoter | This study |
| pRU953 | 1.5-kb EcoRI fragment from pRU942 in pRK415-1 with mctR not under the control of the lac promoter | This study |
| pRU974 | 2.7-kb EcoRI fragment from pRU936 in pRK415-1 with mctSR not under the control of the lac promoter | This study |
| pRU971 | 1.8-kb PCR product (primers P353, P355, P357 and P358; aapJ promoter-gltP fusion) cloned into pCR2.1-TOPO | This study |
| pRU976 | 1.8-kb XbaI/HindIII fragment from pRU971 (aapJ promoter-gltP fusion) in pJP2 | This study |
| pRU980 | 1.8-kb PCR product (primers P354, P356, P357 and P359; aapJ promoter-gltS fusion) cloned into pCR2.1-TOPO | This study |
| pRU986 | 1.8-kb XbaI/HindIII fragment from pRU976 (aapJ promoter-gltS fusion) in pJP2 | This study |