TABLE 1.
Strains, plasmids, and primers used in this study
| Bacterial strain, plasmid, or primer | Genetic characteristics | Antibiotic resistance | Source or reference |
|---|---|---|---|
| S. meliloti | |||
| Rm1021 | Wild-type strain SU47 | Sm | Ausubel |
| CS6000 | Rm1021 ΔcbrA | Sm | (16) |
| CS7001 | CS6000 divLQ362P | Sm | This study |
| HABsup1A | CS6000 divLV602M | Sm | This study |
| HABsup1B | CS6000 divLF142S | Sm | This study |
| HABsup2 | CS6000 divLL454I | Sm | This study |
| HABsup3 | CS6000 divLF142S | Sm | This study |
| HABsup5 | CS6000 cckAA373S | Sm | This study |
| HABsup6 | CS6000 cckAA373S | Sm | This study |
| HABsup7 | CS6000 cckAA373S | Sm | This study |
| HABsup9 | CS6000 divLV787M | Sm | This study |
| JP005 | CS6000 divLV602M Tn5-divL | Sm Nm | This study |
| JP006 | CS6000 divLL454I Tn5-divL | Sm Nm | This study |
| JP007 | CS6000 divLF142S Tn5-divL | Sm Nm | This study |
| JP008 | CS6000 divLV787M Tn5-divL | Sm Nm | This study |
| JP010 | CS6000 divLQ362P Tn5-divL | Sm Nm | This study |
| JP011 | Rm1021 Tn5-divL | Sm Nm | This study |
| JP012 | CS6000 Tn5-divL | Sm Nm | This study |
| JP013 | Rm1021 divLQ362P Tn5-divL | Sm Nm | This study |
| JP014 | Rm1021 divLF142S Tn5-divL | Sm Nm | This study |
| JP015 | Rm1021 divLL454I Tn5-divL | Sm Nm | This study |
| JP016 | Rm1021 divLV602M Tn5-divL | Sm Nm | This study |
| JP017 | Rm1021 divLV787M Tn5-divL | Sm Nm | This study |
| JP001 | CS6000 cckAA373S Tn5-cckA | Sm Nm | This study |
| JP002 | Rm1021 Tn5-cckA | Sm Nm | This study |
| JP003 | Rm1021 cckAA373S Tn5-cckA | Sm Nm | This study |
| JP004 | CS6000 Tn5-cckA | Sm Nm | This study |
| HAB2067 | JP017 pBBR1-MCS-3 | Sm Nm Tc | This study |
| HAB2069 | JP017 pHAB3059 | Sm Nm Tc | This study |
| HAB2031 | Rm1021 pBBR1-MCS-3 | Sm Tc | This study |
| HAB2028 | Rm1021 pHAB3057 | Sm Tc | This study |
| HAB2050 | Rm1021 pHAB3059 | Sm Tc | This study |
| HAB2022 | CS6000 pHAB3059 | Sm Tc | This study |
| HAB2035 | CS6000 pBBR1-MCS-3 | Sm Tc | This study |
| HAB2039 | CS6000 pHAB3057 | Sm Tc | This study |
| HAB2057 | HABsup3 pBBR1-MCS-3 | Sm Tc | This study |
| HAB2052 | HABsup3 pHAB3059 | Sm Tc | This study |
| HAB2045 | CS7001 pHAB3059 | Sm Tc | This study |
| HAB2058 | CS7001 pBBR1-MCS-3 | Sm Tc | This study |
| HAB2037 | HABsup2 pBBR1-MCS-3 | Sm Tc | This study |
| HAB2048 | HABsup2 pHAB3059 | Sm Tc | This study |
| HAB2043 | HABsup1A pBBR1-MCS-3 | Sm Tc | This study |
| HAB2054 | HABsup1A pHAB3059 | Sm Tc | This study |
| HAB2055 | HABsup9 pHAB3059 | Sm Tc | This study |
| HAB2059 | HABsup9 pBBR1-MCS-3 | Sm Tc | This study |
| HAB2056 | HABsup6 pHAB3057 | Sm Tc | This study |
| HAB2060 | HABsup6 pBBR1-MCS-3 | Sm Tc | This study |
| HAB2070 | JP003 pBBR1-MCS-3 | Sm Nm Tc | This study |
| HAB2074 | JP003 pHAB3057 | Sm Nm Tc | This study |
| MB672 | Rm1021 pMB697 | Sm Sp | (20) |
| HAB7010 | Rm1021 pMB697 | Sm Sp | This study |
| HAB7011 | HABsup9 pMB697 | Sm Sp | This study |
| HAB7012 | HABsup6 pMB697 | Sm Sp | This study |
| HAB7013 | HABsup1A pMB697 | Sm Sp | This study |
| HAB7024 | CS6000 pMB697 | Sm Sp | This study |
| HAB7025 | HABsup3 pMB697 | Sm Sp | This study |
| HAB7026 | JP009 pMB697 | Sm Sp | This study |
| HAB7027 | HABsup2 pMB697 | Sm Sp | This study |
| HAB8001 | Rm1021 pJP1001 | Sm Nm | This study |
| HAB8002 | CS6000 pJP1001 | Sm Nm | This study |
| HAB8003 | CS7001 pJP1001 | Sm Nm | This study |
| HAB8004 | HABsup1A pJP1001 | Sm Nm | This study |
| HAB8005 | HABsup1B pJP1001 | Sm Nm | This study |
| HAB8006 | HABsup2 pJP1001 | Sm Nm | This study |
| HAB8007 | HABsup3 pJP1001 | Sm Nm | This study |
| HAB8008 | HABsup5 pJP1001 | Sm Nm | This study |
| HAB8009 | HABsup6 pJP1001 | Sm Nm | This study |
| HAB8010 | HABsup7 pJP1001 | Sm Nm | This study |
| HAB8011 | HABsup9 pJP1001 | Sm Nm | This study |
| RH005 | Rm1021 pK19ms::dnaN-mCherry | Sm Gm | This study |
| RH006 | CS6000 pK19ms::dnaN-mCherry | Sm Gm | This study |
| E. coli | |||
| MT616 | MM294 pRK600 | Cm | Finan |
| DH5α | endA1 hsdR17 supE44 thi-1 recA1 gyrA relA1Δ(lacZYA-argG) | BRL Corp. | |
| BL21 | (DE3) pLysS ompT hsdSB (rB− mB−) gal dcm | Cm | Invitrogen |
| Plasmids | |||
| pBBR1-MCS-3 | Low copy vector | Tc | (34) |
| pHAB3057 | pBBR1-MCS-3::cckAWT | Tc | This study |
| pHAB3059 | pBBR1-MCS-3::divLWT | Tc | This study |
| pSF-OXB20-HN2-V5-TEV | High Copy Expression vector, N-terminal V5 | Kn | Oxford Genetics |
| pHAB4011 | pSF-OXB20-HN2-V5-TEV:: cckAΔTM | Kn | This study |
| pHAB4038 | pSF-OXB20-HN2-V5-TEV:: cckAΔTM_A373S | Kn | This study |
| pLAFR1 | Low copy vector | Tc | (35) |
| pLAFR2070 | cbrAWT complementation vector | Tc | (18) |
| pVO155 | Suicide vector with uidA cassette | Nm | (36) |
| pMB697 | pVO345::SMc00888-uidA | Sp | (18) |
| pJB1001 | pVO155::ftsk2-uidA | Nm | This study |
| pK19ms::dnaN-mCherry | pK19mobsacB with dnaN-mCherry translation fusion | Kn Gm | (12) |
| Primers | Sequence 5′–3′ | ||
| cat1 | AACTCACCCAGGGATTGGCT | (16) | |
| cat2 | ACCAGACCGTTCAGCTGGATA | (16) | |
| HAB0002 | GCAGGCATTCAGTCGCT | This study | |
| HAB0003 | GGTACGCTTGATTCATCG | This study | |
| HAB0004 | ATGTCCTGACGGCCATC | This study | |
| HAB0005 | ACGACCTTCCGCATCCT | This study | |
| HAB0007 | AGCGACTGAATGCCTGC | This study | |
| HAB0008 | CGATGAATCAAGCGTACC | This study | |
| HAB0009 | GATGGCCGTCAGGACAT | This study | |
| HAB0010 | AGGATGCGGAAGGTCGT | This study | |
| HAB0011 | CAAACCGGAAACGGATTGGG | This study | |
| HAB0012 | TGAATGCGGGGTTCGGTTG | This study | |
| HAB0015 | GACTAGTCTGAACCAGGGTGAAGGTTGG | This study | |
| HAB0016 | GGGGTACCCCCGAGCTCTCGGAGGAACAGG | This study | |
| HAB0017 | CGAGCTCGCGGATACTGCTCATGAAAGG | This study | |
| HAB0018 | GACTAGTCAAGGGAGAGGAGGGCTTGTT | This study | |
| HAB0021 | GAATTCGATCGAGGTGATGCCGCAATCG | This study | |
| HAB0022 | GGATCCTCAGCTGTCGAGCATCTCGCG | This study | |
| CTP050 | ATAAGGATCCTGAACCAGGGTGAAGGTTGG | This study | |
| CTP049 | ATTACATATGGGCAAACAGACGGAGACAGA | This study | |
| CTP019 | AGGCCAGCTCGAAAAGGCGAT | This study | |
| CTP020 | AATCAGCGGCTGCAGTTCTACAAT | This study | |
| CTP021 | ATCCTCGACTACGCGGTCATT | This study | |
| CTP022 | TTCAACCGCTTCGAAAGCTAC | This study | |
| CTP023 | TGTCGGGTGCATCGGTTCGAA | This study | |
| CTP024 | GATGCCCCAGATCGCCCGGAA | This study | |
| CTP025 | AAACCTTGTCGCGGAAAGGCGTCT | This study | |
| CTP026 | TGTTCTTGTCGGCAATGAGAG | This study | |
| HAB0023 | GCAAAGAGGCGGGAGAGATT | This study | |
| HAB0024 | TCATCCACAGGAACCGCAAA | This study | |
| HAB0025 | CAGATGTGAACAAGTCGCCG | This study | |
| HAB0026 | TCCGCGGTCTGTTGATAAGG | This study | |
| HAB0027 | GATTTCTGGAGTTCTTGGGCG | This study | |
| HAB0028 | AGGCTTTTCACGTCGGGAG | This study | |
| HAB0029 | TTCTGGAGTTCTTGGGCGGT | This study | |
| HAB0030 | GCGACAAGGCTTTTCACGTC | This study | |
| HAB0031 | GGAGCGCTCGCAATTCATAC | This study | |
| HAB0032 | AGACCCTGCGGATCATCTCT | This study | |
| HAB0033 | GCAGGAGACGATGGATGGAA | This study | |
| HAB0034 | GAAACGCTGATCGCTCTCCA | This study | |
| HAB0035 | CGAGCTCCTTCGTGACTCTG | This study | |
| HAB0036 | AGAGAGGCGCAGACAATTCC | This study | |
| HAB0037 | CAGCCTCGCTTTCATGGAGA | This study | |
| HAB0038 | TCGGTAGAGGTGAACTTGCG | This study | |
| HAB0039 | GACGTGATCTGCAAGTCCCT | This study | |
| HAB0040 | CGGACCAATCTGCATCAGGA | This study | |
| HAB0041 | GAGCGGCACAAACAGGAATC | This study | |
| HAB0042 | CCTTGTTGGCCGGTTCAATG | This study | |
| ftsK2-1 | ACTAGTCCACTTCTGAAACGACAGC | This study | |
| ftsK2-2 | CTCGAGCGACGAACGCTGTATCAC | This study | |