TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Relevant genotype or characteristics | Source or reference |
|---|---|---|
| Strains | ||
| E. coli DH5α | F−recA lacZΔM15 | BRL |
| R. solanacearum | ||
| GMI1000 | Wild-type strain | 8 |
| GMI1402 | hrcS::Tn5-B20 mutant | 4 |
| GMI1410 | hrpY::Tn5-B20 mutant | 4 |
| GMI1475 | hrpB::Tn5-B20 mutant | 4 |
| GMI1551 | PopA::ΩSp/Sm | 5 |
| GMI1575 | prhA::Ω mutant | 50 |
| GMI1663 | popF1::ΩSp/Sm mutant | This study |
| GMI1664 | popF2::ΩCm mutant | This study |
| GMI1665 | popF2:: aprA (Gm) mutant | This study |
| GMI1666 | popF1::ΩSp/SmpopF2::ΩCm double mutant | This study |
| GMI1667 | popF1::ΩSp/SmpopF2::aprA (Gmr) double mutant | This study |
| GMI1668 | popF2-His6, Spr/Smr | This study |
| GMI1669 | popF2-His6hrcS::Tn5-B20, Spr/Smr Kmr | This study |
| GMI1670 | popF2-His6 (Sp/Sm) hrpY::Tn5-B20, Spr/Smr Kmr | This study |
| X. campestris pv. campestris 8004 | Wild type, Rifr | 73 |
| Plasmids | ||
| pBluescript II KS(−) | Cloning vector, Ampr | Stratagene |
| pGEM-T | Cloning vector, Ampr | Promega |
| pHP45Ω Sp/Sm | Spr, Smr, Ω cassette | 57 |
| pHP45ΩCm | Cmr, Ω cassette | 57 |
| pLAFR6 | Tcr | 35 |
| pMS107 | pIC20H containing the cyaA gene | 67 |
| pUC1318 Apra/Gm | Gmr, aprA cassette | 9 |
| pAM5 | pLAFR3 with a 2-kb insert containing hrpB, Tcr | 28 |
| pCL1 | pGEM T with a 2.5-kb insert containing popF2, Ampr | This study |
| pCL3 | pGEM-T with a 1.5-kb fragment containing popF1 N-terminal DNA region, Ampr | This study |
| pMG6 | pBluescript KS(−) with a 1-kb XbaI HindIII fragment containing popF2 C-terminal DNA region, Ampr | This study |
| pCL86 | pCZ205 with a 712-bp HindIII XbaI fragment carrying popF2 promoter, Ampr Gmr | This study |
| pCL94 | pLAFR6 carrying a 4.7-kb fragment carrying popF2 promoter and promoterless lacZ, Tcr Gmr | This study |
| pCZ205 | pUC19 carrying lacZ in KpnI-XbaI, Ampr Gmr | 46 |
| pCZ367 | pUC18-derived vector used for insertional mutagenesis, Ampr Gmr | 21 |
| pCZ406 | pCZ205 with a 600-bp HindIII XbaI fragment carrying popF1 promoter, Ampr Gmr | This study |
| pCZ408 | pLAFR6 carrying a 4.6-kb fragment carrying popF1 promoter and promoterless lacZ, Tcr Gmr | This study |
| pHrpF | pLAFR6 with a 3.5-kb HindIII-EcoRI fragment containing hrpFXcc, Tcr | This study |
| pMG7 | pMG6 with a 1-kb HindIII XhoI fragment containing popF2 C-terminal DNA region, Ampr | This study |
| pMDI | pGEM-T with a 3.5-kb HindIII-EcoRI fragment containing HrpFXcc, Ampr | This study |
| pPopF1 | pLAFR6 with a 2.5-kb HindIII-KpnI fragment carrying popF1 | This study |
| pPopF2 | pLAFR6 with a 2.5-kb HindIII-KpnI fragment carrying popF1 | This study |
| pSC154 | pET-26b(+)-derived vector with the cyaA′ gene from pMS107 used for translational fusion constructs; the T7 promoter is replaced by a ptac promoter, Kmr | 21 |
| pSC163 | PLAFR6 carrying a translational fusion between avrA1-99 and cyaA′ | 19 |