TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Characteristics | Reference and/or source |
|---|---|---|
| Strains | ||
| E. coli DH5α | supE44 ΔlacU169(φ80lacZΔM15) hsdR17 recA1 endA1 gyrA96 thi-1 relA1, Nalr | 35; Life Technologies |
| E. coli DB3.1 | F gyrA462 endA1 Δ(sr1-recA) mcrB mrr hsdS20 (rB−, mB−) supE44 ara-14 galK2 lacY1 proA2 rpsL20 (Smr) xyl-5 λ−leu mtl-1 | Invitrogen |
| P. syringae pv. tomato DC3000 | Wild type; spontaneous Rifr | 23 |
| P. syringae pv. syringae B728A | Wild type; spontaneous Rifr | 37 |
| P. syringae pv. tomato UNL137 | DC3000 hopO1-1 operon deletion mutant, Rifr Spr | This work |
| Plasmids | ||
| pBluescript-II KS+ | Cloning vector, Apr | Stratagene |
| pML123 | Broad-host-range cloning vector, Gmr Kmr | 48 |
| pBBR1MCS1 | Broad-host-range vector, Cmr | 47 |
| pRK415 | Broad-host-range vector, unstable in absence of selection, Tcr | 45 |
| pEG202 | Yeast two-hybrid vector, creates fusion to LexA DNA binding domain, Apr | 32 |
| pJG4-5 | Yeast two-hybrid vector, creates fusion to B42 activation domain, Apr | 32 |
| pENTR/D-TOPO | Gateway system donor vector, Kmr | Invitrogen |
| pCPP2318 | pCPP30 derivative carrying blaM lacking signal peptide sequences, Tcr | 19 |
| pCPP3234 | Gateway destination vector containing the adenylate cyclase (cyaA) gene for C-terminal fusions, Spr/Smr Cmr | 62 |
| pLN142 | pML123 derivative carrying hopO1-1-flag, Gmr | 59 |
| pLN241 | pJG4-5 derivative carrying hopT1-1, Apr | This work |
| pLN248 | pJG4-5 derivative carrying hopO1-1, Apr | This work |
| pLN249 | pEG202 derivative carrying shcO1, Apr | This work |
| pLN250 | pJG4-5 derivative carrying shcO1, Apr | This work |
| pLN256 | pML123 derivative carrying hopT1-1-flag, Gmr | This work |
| pLN314 | pBluescript-II KS+ derivative containing 1.6 kb DNA region upstream of shcO1, omega cassette, and 2.0 kb DNA region downstream of hopT1-1, Apr Spr | This work |
| pLN366 | pENTR/D-TOPO derivative carrying hopS2, Kmr | This work |
| pLN367 | pENTR/D-TOPO derivative carrying hopS1, Kmr | This work |
| pLN370 | pRK415 derivative containing upstream and downstream flanking sequences of hopO1-1 operon with omega cassette in between, Tcr Spr | This work |
| pLN411 | pML123 derivative carrying shcO1/hopO1-1-flag, Gmr | This work |
| pLN428 | pJG4-5 derivative carrying hopO1-2, Apr | This work |
| pLN454 | pBBR1MCS5 derivative carrying hopO1-1 operon, Gmr | This work |
| pLN521 | pML123 derivative carrying hopO1-2-flag, Gmr | This work |
| pLN567 | pML123 derivative carrying hopT1-2-flag, Gmr | This work |
| pLN568 | pEG202 derivative carrying shcS1, Apr | This work |
| pLN622 | pENTR/D-TOPO derivative carrying shcS2lhopS2, Kmr | This work |
| pLN625 | pJG4-5 derivative carrying hopO1-1 fragment corresponding to HopO1-11-140, Apr | This work |
| pLN627 | pJG4-5 derivative carrying hopO1-1 fragment corresponding to HopO1-1141-283, Apr | This work |
| pLN705 | pBBR1MCS1 Gateway destination vector containing a hemagglutinin tag for C-terminal fusions, Cmr | This work |
| pLN716 | pJG4-5 derivative carrying hopO1-1 fragment corresponding to HopO1-1190-283, Apr | This work |
| pLN750 | pJG4-5 derivative carrying shcS1, Apr | This work |
| pLN752 | pJG4-5 derivative carrying hopT1-2, Apr | This work |
| pLN753 | pJG4-5 derivative carrying hopS1, Apr | This work |
| pLN786 | pENTR/D-TOPO derivative carrying shcO1, Kmr | This work |
| pLN814 | pENTR/D-TOPO derivative carrying hopO1-1, Kmr | This work |
| pLN898 | Gateway construct made by recombining pLN705 and pLN786, Cmr | This work |
| pLN966 | pEG202 derivative carrying shcS2, Apr | This work |
| pLN967 | pJG4-5 derivative carrying hopS2, Apr | This work |
| pLN980 | pJG4-5 derivative carrying shcS2, Apr | This work |
| pLN998 | pJG4-5 derivative carrying hopO1-1 fragment corresponding to HopO1-1111-175, Apr | This work |
| pLN999 | pJG4-5 derivative carrying hopO1-1 fragment corresponding to HopO1-171-189, Apr | This work |
| pLN1003 | pENTR/D-TOPO derivative carrying shcS1/hopS1, Kmr | This work |
| pLN1005 | Gateway construct made by recombining pCPP3234 and pLN1003, Spr | This work |
| pLN1006 | Gateway construct made by recombining pCPP3234 and pLN367, Spr | This work |
| pLN1018 | pML123 derivative carrying shcS2-ha/hopO1-1-flag, Gmr | This work |
| pLN1019 | pML123 derivative carrying shcS1-ha/hopO1-1-flag, Gmr | This work |
| pLN1022 | pJG4-5 derivative carrying hopO1-1 fragment corresponding to HopO1-11-125, Apr | This work |
| pLN1023 | pJG4-5 derivative carrying hopO1-1 fragment corresponding to HopO1-11-110, Apr | This work |
| pLN1027 | pJG4-5 derivative carrying hopO1-1 fragment corresponding to HopO1-1161-283, Apr | This work |
| pLN1063 | pENTR/D-TOPO derivative carrying shcO1/hopO1-1, Kmr | This work |
| pLN1064 | pENTR/D-TOPO derivative carrying shcS2-ha/hopO1-1, Kmr | This work |
| pLN1065 | pENTR/D-TOPO derivative carrying shcS1-ha/hopO1-1, Kmr | This work |
| pLN1074 | Gateway construct made by recombining pCPP3234 and pLN1063, Spr | This work |
| pLN1075 | Gateway construct made by recombining pCPP3234 and pLN1064, Spr | This work |
| pLN1076 | Gateway construct made by recombining pCPP3234 and pLN1065, Spr | This work |
| pLN1077 | Gateway construct made by recombining pCPP3234 and pLN814, Spr | This work |
| pLN1090 | pENTR/D-TOPO derivative carrying shcS2/hopS2 containing an S4I site-directed mutation in HopS2, Kmr | This work |
| pLN1138 | Gateway construct made by recombining pCPP3234 and pLN622, Spr | This work |
| pLN1140 | Gateway construct made by recombining pCPP3234 and pLN1090, Spr | This work |
| pLN1233 | Gateway construct made by recombining pCPP3234 and pLN366, Spr | This work |
| pLN1234 | pENTR/D-TOPO derivative carrying shcS2/hopS2 from P. syringae pv. tomato PT23 | This work |
| pLN1235 | pENTR/D-TOPO derivative carrying shcS2/hopS2 from P. syringae pv. maculicola 84-67 | This work |
| pLN1236 | pENTR/D-TOPO derivative carrying shcS2/hopS2 from P. syringae pv. maculicola NCPPB1886 | This work |
| pLN1243 | Gateway construct made by recombining pCPP3234 and pLN1245, Spr | This work |
| pLN1245 | pENTR/D-TOPO derivative carrying hopS2 containing an S4I site-directed mutation in HopS2, Kmr | This work |
| pLN1460 | pJG4-5 derivative carrying hopO1-1 fragment corresponding to HopO1-11-70, Apr | This work |