TABLE 1.
Strains and plasmids used in this work
| Strain or plasmid | Relevant characteristics | Source or reference |
|---|---|---|
| Strain | ||
| R. etli | ||
| CFN42 | Wild type, Nalr | 50 |
| CE3 | CFN42 derivative, Strr | 50 |
| CFNX101 | CE3 derivative, recA::Sp | 39 |
| CFNX693 | CE3 derivative, recF::loxPSp | This work |
| CFNX694 | CE3 derivative, addA::loxPSp | This work |
| CFNX695 | CE3 derivative with a deletion-substitution of addAaddB by ΔaddAB::loxPSp | This work |
| CFNX696 | CFNX693 derivative, but recF::loxP | This work |
| CFNX697 | CFNX696 derivative, but addA::loxPSp | This work |
| E. coli | ||
| DH5α | recA1 | Stratagene |
| S17.1 | recA13, mobilization host | 56 |
| Plasmid | ||
| pBluescript II SK(+) | Cbr, cloning vector | Stratagene |
| pSUP202 | Cmr Tcr Cbr, suicide vector | 56 |
| pKOK4 | Cmr Tcr Cbr, suicide vector | 23 |
| pJQ200SK+ | GmrsacB, suicide vector | 47 |
| pBX404-7 | Tcr 5′ del neo/3′ del neo Kms plasmid recombination system | 65 |
| pBBRMCS5 | Gmr, broad-host-range vector | 28 |
| pJMS2 | Spr, source of loxPSp cassette | 40 |
| pJMS08 | vector containing the loxP site-specific recombinase Cre | Martínez-Salazar et al., unpublished |
| pLGsym56 | pSUP202 derivative harboring a 6.0-kb BamHI fragment from R. etli pSym | 19 |
| pLG506 | pSUP202 derivative containing 2.0-kb HindIII ΩKm and MCS from pBluescript SK(+) into BgIII | Girard et al., unpublished |
| pRK2013 | Kmr, helper vector | 56 |
| c1343 | pLAFR1 cosmid harboring R. etli recF region | This work |
| c2486 | pLAFR1 cosmid harboring R. etli addAB region | This work |
| pJZC1 | pKOK4 derivative harboring the 3.0-kb EcoRI fragment of R. etli recF gene | This work |
| pJZC2 | pJZC1 derivative containing the R. etli recF::loxPSp allele | This work |
| pJZC3 | pSK derivative harboring a 6-kb EcoRI fragment of R. etli addA gene | This work |
| pJZC4 | pSK derivative harboring a 1.5-kb EcoRI PCR product with 225 bp upstream and 1,188 bp of R. etli addA gene | This work |
| pJZC5 | pJZC4 derivative harboring the R. etli addA::loxPSp allele | This work |
| pJZC6 | pJQ200SK derivative harboring the 3.5-kb EcoRI addA::loxPSp from pJZC5 into MCS | This work |
| pJZC7 | pJQ200SK derivative containing the 3.3-kb EcoRI-SalI 5′ del neo/3′ del neo Kms from pBX404-7 into MCS | This work |
| pJZC8 | pJZC7 derivative containing the 6.0-kb BamHI fragment from pLGsym56 into MCS | This work |
| pJZC9 | pBBMCS5 derivative harboring the addA gene in the same orientation to lacZ promoter | This work |
| pJZC10 | pBBMCS5 derivative harboring the addA gene in the opposite orientation to lacZ promoter | This work |
| pJZC11 | pBBMCS5 derivative harboring the addAB genes in the same orientation to lacZ promoter | This work |
| pJZC12 | pSK derivative containing the 1.2-kb SalI from pJZC3, corresponding to 585 bp of 3′ end of addA and 672 bp downstream (trxA and folC) | This work |
| pJZC13 | pJZC12 derivative harboring the 2.0-kb SalI loxPSp into the SalI site located at aa 988 of addA | This work |
| pJZC14 | pSK derivative harboring the 1.2-kb EcoRI-XhoI, corresponding to aa 63 to 469 of addB | This work |
| pJZC15 | pJZC13 derivative harboring 1.2-kb EcoRI-XhoI from pJZC14 into EcoRI-SalI, ΔaddAB::loxPSp | This work |
| pJZC16 | pLG506 derivative harboring the 4.4-kb EcoRI-XhoI from pJZC15 (ΔaddAB::loxPSp) | This work |