Table 1.
Bacterial strains, plasmids, and primers used in this work
| Strain, plasmid, or primer | Relevant characteristics | Source or reference |
|---|---|---|
| Strains | ||
| 8004 | Wild type, Rifr | 32 |
| ΔhrpX strain | hrpX deletion mutation in strain 8004 | This study |
| Plasmids | ||
| pLAFR6 | pLAFR1 with rho-independent terminator and pUC18 polylinker, Tetr | 11 |
| pFR421 | pLAFR6(PpipXcc-gusA) | 19 |
| pHM1 | Broad-host-range cosmid vector, pSa ori, Spr | 19 |
| pHM1-hrpX-his | pHM1 (hrpX-his) | This study |
| pMAL-p2X | Tac promoter, expression vector, Ampr | New England BioLabs |
| pMAL-p2x/hrpX | pMAL-p2X(hrpX-his) | This study |
| pGEM-T | Cloning vector | Promega Co. |
| pK18mobsacB | Suicide plasmid in X. campestris pv. campestris, Kmr | 29 |
| Primers | ||
| hrpXhisF | 5′cccaagcttATGATCCTTTCGACCTACTTCGC3′a | |
| hrpXhisR | 5′ccggaattctcagtggtggtggtggtggtgGCGTTGCAGGGTTTCCAT3′ | |
| jxhrpXFwd | 5′ttccatatgATGATCCTTTCGACCTACTTCGCA3′ | |
| jxhrpXRev | 5′ccgctcgagGCGTTGCAGGGTTTCCATCGG3′ | |
| hrpX del F1 | 5′gatatcATGATCCTTTCGACCTACTTGA3′ | |
| hrpX del R1 | 5′gtcgacTGGAAGTGGGTCAGCGCCTT3′ | |
| hrpX del F2 | 5′gtcgacCGCCCTGGGGGCTGTGCAA3′ | |
| hrpX del R2 | 5′aagcttTTAGCGTTGCAGGGTTTCCATCG3′ | |
| hrpX outup | 5′GCTCACCGCTGCCCTGCATTGCTGC3′ | |
| hrpX outdown | 5′TACAATCGTTTGCGCCCACCACAAC3′ | |
| box45F | 5′ACGGTGTCGCAATTCGCGCGTTTCGCAATTGCCAACCGGTGTCAT3′ | |
| box45R | 5′ATGACACCGGTTGGCAATTGCGAAACGCGCGAATTGCGA3′ | |
| pip-PF | 5′GTCGAATTCGAAGGCTCAGTTGGTCGGGTTTG3′ | |
| pip-PR | 5′CTCGGTGCACTTCATGACCTGC3′ | |
| pipF1 | 5′GAAGGCTCAGTTGGTCGGGTT3′ | |
| pipR1 | 5′GGCAATTGCGAAACGCGCGAA3′ | |
| pipR2 | 5′GACCTGCGCCCACTTACGG3′ |
a
The lowercase letters in primer sequences indicate the restriction enzyme recognition sites and fusion tag sequences.