TABLE 1.
Strains and plasmids
| Designation | Main feature | Characteristics | Reference and/ or source |
|---|---|---|---|
| Bacterial strains | |||
| E. coli DH5α | General use | supE44 ΔlacU169(φ80lacZΔM15) hsdR17 recA1 endA1 gyrA96 thi-1 relA1 Nair | 33; Life Technologies |
| P. syringae pv. tomato | |||
| DC3000 | Wild-type strain | spontaneous Rifr | 18 |
| DC3000 hrcC mutant | TTSS defective | DC3000 hrcC mutant, Rifr Cmr | 70 |
| UNL140 | hrpJ mutant | DC3000 nonpolar hrpJ deletion, Rifr Kmr | This work |
| Plasmids | |||
| pAvrRpt2-600 | Broad-host-range construct encoding AvrRpt2 | pDSK600 derivative, Spr/Smr | 62 |
| pBBR1MCS1/5 | Broad-host-range vector | Cmr (MCS1) or Gmr (MCS5) | 47 |
| pBluescript-II KS(−) | Cloning vector | Apr | Stratagene |
| pCPP2308 | Broad-host-range construct encoding HrpL | pML122 derivative, Gmr | Alan Collmer |
| pCPP2318 | Broad-host-range construct encoding mature β-lactamase | pCPP30 derivative carrying blaM lacking signal peptide sequences, Tcr | 14 |
| pCPP2330 | Broad-host-range construct encoding AvrB1-FLAG | pML123 derivative, Gmr | 28 |
| pCPP2988 | Construct carrying nptII | pBluescript II KS(−) vector carrying 1.5-kb HindIII-SalI fragment with nptII lacking transcriptional terminator, Apr Kmr | 1 |
| pCPP3234 | Gateway destination vector for CyaA fusions | Spr Smr | 64 |
| pCPP5040 | Broad-host-range Gateway destination vector for HA fusions | pML123 derivative, Gmr | 42 |
| pENTR/D-TOPO | Gateway entry vector | Kmr | Invitrogen |
| pLN307 | avrPto1 entry construct | Kmr | This work |
| pLN323 | avrPtoB entry construct | Kmr | 42 |
| pLN375 | hrpJ entry construct | Kmr | This work |
| pLN420 | hopB1 entry construct | pENTR/D-TOPO derivative, Kmr | 60 |
| pLN421 | Broad-host-range construct encoding HopB1-CyaA | pCPP3234 derivative obtained by recombination with pLN420, Spr/Smr | 60 |
| pLN426 | Broad-host-range construct encoding HrpJ-CyaA | pML123 derivative obtained by recombination with pLN375, Gmr | This work |
| pLN677 | Broad-host-range Gateway destination vector for HA fusions | pBBR1MCS-5 derivative, Gmr | 60 |
| pLN705 | Broad-host-range Gateway destination vector for HA fusions | pBBR1MCS-1 derivative, Cmr | 31 |
| pLN726 | Broad-host-range construct encoding HrpJ-HA | pLN677 derivative obtained by recombination with pLN375, Gmr | This work |
| pLN736 | Broad-host-range construct encoding HrpJ-HA | pLN705 derivative obtained by recombination with pLN375, Cmr | This work |
| pLN820 | avrB1 entry construct | pENTR/D-TOPO derivative, Kmr | This work |
| pLN918 | Broad-host-range construct encoding AvrB1-CyaA | pCPP3234 derivative obtained by recombination with pLN820, Spr/Smr | This work |
| pLN1979 | Broad-host-range construct encoding AvrPtoB-CyaA | pCPP3234 derivative obtained by recombination with pLN323, Spr/Smr | This work |
| pLN1985 | Broad-host-range construct encoding AvrPto1-CyaA | pCPP3234 derivative obtained by recombination with pLN307, Spr/Smr | This work |
| pLN2043 | Construct carrying cyaA | pBluescript II KS(−) derivative containing cyaA, Apr | This work |
| pLN2179 | Construct carrying DNA upstream of hrpJ adjacent to an nptII cassette | pBluescript II KS(−) derivative, Apr Kmr | This work |
| pLN2180 | Broad-host-range hrpJ deletion construct | pBluescript II KS(−) derivative, Apr Kmr | This work |
| pLN2190 | Construct carrying cyaA with flanking Gateway recombination sites | pBluescript II KS(−) derivative, Apr Cmr | This work |
| pLN2193 | Gateway destination vector for CyaA fusions | pML123 derivative, Gmr Cmr | This work |
| pLN2234 | Broad-host-range construct encoding HrpJ-CyaA | pLN2193 derivative obtained by recombination with pLN375, Gmr | This work |
| pLN2302 | hrpJ deletion construct | pRK415 derivative carrying insert from pLN2180, Tcr | This work |
| pML122/23 | Broad-host-range vector | Gmr | 49 |
| pRK415 | Broad-host-range vector | Tcr | 45 |