TABLE 1.
Bacterial strains and plasmids used
| Strain or plasmid | Description | Source or reference |
|---|---|---|
| Strains | ||
| R. leguminosarum | ||
| 3841 | Strr derivative of R. leguminosarum biovar viciae strain 300 | 10, 14 |
| 3855 | Strr derivative of R. leguminosarum biovar viciae | 38 |
| CR534 | Strain 3855 dctA::Tn5 | 38 |
| CR535 | Strain 3855 dctB::Tn5 | 38 |
| CR538 | Strain 3855 dctD::Tn5 | 38 |
| RU150 | Strain 3841 dctA101 | This work |
| RU152-1 | Second-site revertant of RU150 | This work |
| RU152-14 | Second-site revertant of RU150 | This work |
| RU152-22 | Second-site revertant of RU150 | This work |
| RU436 | Strain 3841 dctA::Tn5 | 29 |
| RU437 | Strain 3841 dctA::Tn5 | 29 |
| RU711 | Strain 3841 ΔdctD::Ω Strr Spcr | 34 |
| RU714 | Strain 3841 ΔdctABD::Ω Strr Spcr | 34 |
| RU720 | Strain RU150 dctA101 ΔdctD::Ω Strr Spcr Kmr | This work |
| RU727 | Strain 3841 dctA::Ω Strr Spcr | 34 |
| RU730 | Strain 3841 dctB::Ω Strr Spcr | 33 |
| RU865 | Strain 3841 ΔdctBD::Ω Strr Spcr | 33 |
| RU873 | Strain RU150 dctA101 ΔdctBD::Ω Strr Spcr Kmr | This work |
| RU875 | Strain RU150 dctA101 dcB::Ω Strr Spcr Kmr | This work |
| RU929 | Strain 3841 ntrC::Ω Strr Spcr | 33 |
| RU938 | Strain RU150 dctAB::Tn5 dcA::Ω Strr Spcr Kmr | This work |
| E. coli S17-1 | pro hsdR recA [RP4-2(Tc::Mu) (Km::Tn7)]; RP4 integrated into its chromosome | 42 |
| Plasmids | ||
| Bluescript II SK− | Phagemid; f1− origin of replication; ColE1 replicon; SK polylinker; 2.96 kb; standard cloning vector; Ampr | Stratagene Ltd. |
| pGEMT | T overhang vector for PCR product cloning; Ampr | Promega |
| pMP220 | IncP broad-host-range mobilizable promoter probe vector employing E. coli lacZ as reporter gene; 10.5 kb; Tcr | 43 |
| pJQ200KS | Narrow host range (p15A ori); mob sacB; 5.4 kb; Gmr | 31 |
| pHP45Ω | 4.3-kb plasmid containing 2.0-kb Ω interposon encoding the gene for Spr with transcription and translation termination signals at both ends; Ampr Spcr | 30 |
| pIJ1848 | Cosmid containing dctABD from R. leguminosarum 8002; Tcr | 25 |
| pIJ1969 | pIJ1848 dctB::Tn5 | 25 |
| pIJ1970 | pIJ1848 dctA::Tn5 | 25 |
| pRU10 | 6.7-kb EcoRI chromosomal Tn5 bearing fragment from strain RU150 cloned in Bluescript SK−; Ampr | This work |
| pRU16 | 0.9-kb dctA-dctB intergenic region from pRU3001 in Bluescript SK−; Ampr | This work |
| pRU47 | 10.4-kb HindIII fragment from pIJ1848 in Bluescript SK−; Ampr | This work |
| pRU69 | 3.0-kb HindIII/EcoRI fragment from pRU150 ligated into pBC SK+; Cmr | This work |
| pRU75 | 1-kb HpaI/HindIII fragment from IS50 cloned in Bluescript SK−; Ampr | This work |
| pRU80 | 2.8-kb internal BglII fragment from Tn5 ligated in Bluescript pBC SK+; Cmr Kmr | This work |
| pRU93 | 6.0-kb EcoRI fragment containing Tn5 cloned from the chromosome of strain RU152-22 into Bluescript SK−; Ampr Kmr | This work |
| pRU103 | 0.9-kb dctA-dctB intergenic region EcoRI from pRU16 in pMP220; dctA-lacZ; Tcr | 33 |
| pRU104 | 0.9-kb dctA-dctB intergenic region EcoRI from pRU16 in pMP220; dctB-lacZ; Tcr | This work |
| pRU105 | dctA101-lacZ fusion in pMP220; Tcr Kmr | This work |
| pRU106 | dctA101B-lacZ fusion in pMP220; Tcr Kmr | This work |
| pRU108 | 3.0-kb EcoRI fragment containing dctA from pRU47 in pMP220; Tcr | This work |
| pRU110 | 0.4-kb inverse PCR fragment from strain RU152-14 cloned in pGEM-T; Ampr | This work |
| pRU123 | 1.5-kb dctA produced by PCR in pGEMT; Ampr | This work |
| pRU150 | pRU47 ClaI digested and religated (6.1 kb); contains dctABD; Ampr | This work |
| pRU155 | pRU123 dctA::Ω; Ampr Spcr | This work |
| pRU158 | ΔdctB::Ω in SK−; Ampr Spcr | This work |
| pRU168 | ΔdctD::Ω in pJQ200KS; Gmr Spcr | This work |
| pRU292 | 0.53-kb PstI fragment from pRU3001 in pMP220; dctD1-lacZ; Tcr | This work |
| pRU303 | 4.6-kb NotI/ApaI fragment in pJQ200KS; ΔdctBD::Ω; Gmr Spcr | This work |
| pRU304 | 4.5-kb NotI/ApaI fragment from pRU347 ligated into pJQ200KS; Gmr Spcr | This work |
| pRU305 | 4.0-kb NotI/ApaI fragment from pRU345 in pJQ200KS; Gmr Spcr | This work |
| pRU345 | 3.1-kb AccIII fragment from pRU10 ligated into pRU155; Ampr Spcr Kmr | This work |
| pRU347 | 6.7-kb EcoRI fragment from pRU10 ligated into pRU158; Ampr Spcr Kmr | This work |
| pRU348 | 3.6-kb BamHI/MluI fragment from pIJ1848 in Bluescript SK−; Ampr | This work |
| pRU352 | dctA101BD-lacZ fusion in pMP220; Tcr Kmr | This work |
| pRU354 | 3.6-kb BamHI/MluI fragment from pIJ1848 in pMP220; dctBD-lacZ; Tcr | This work |
| pRU392 | 2.4-kb EcoRI/XbaI fragment from pIJ1848 in pMP220; dctD2-lacZ; Tcr | This work |
| pRU402 | 0.8-kb PstI/NotI fragment of DNA immediately downstream of dctA in Bluescript pBC SK+; Cmr | This work |
| pTH2A | S. meliloti dctA::TnphoA translational fusion; Tcr | 45 |
| pRU3001 | Cosmid from strain 3841 containing the dct region; Tcr | 29 |