TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Relevant characteristics | Reference or source |
|---|---|---|
| Escherichia coli | ||
| DH5α | 48 | |
| Pseudomonas syringae pv. syringae | ||
| FF5 | Cus; no detectable plasmids, nonmucoid | 28 |
| FF5.31 | Cur Kmr; contains pPSR12, nonmucoid, algL::Tn5 | 44 |
| FF5.32 | Cur Kmr; contains pPSR12, nonmucoid, algR1::Tn5 | 28 |
| Plasmids | ||
| pPSR12 | Cur Smr; 200 kb, confers constitutive alginate production to P. syringae pv. syringae FF5 | 28 |
| pSK2 | Tcr; contains alginate biosynthetic cluster from P. syringae pv. syringae FF5 in pRK7813 | 44 |
| pRK2013 | Kmr; helper plasmid | 15 |
| pRK415 | Tcr; RK2-derived cloning vector | 26 |
| pRK7813 | Tcr; cosmid vector | 23 |
| pCP13 | Tcr; cosmid vector | 10 |
| pBluescript SK(+) | Apr; ColE1 origin, cloning vehicle | Stratagene |
| pRG960sd | Smr Spr; contains promoterless uidA with start codon and Shine-Dalgarno sequence | 58 |
| pBBR1MCS | Cmr; 4.7-kb broad-host-range cloning vector | 30 |
| pBBR.Gus | Cmr; 6.6-kb promoter probe vector containing uidA in pBBR1MCS | 43 |
| pCR2.1 | Apr Kmr; 3.9-kb cloning vector | Invitrogen |
| pADP | Apr Kmr; contains PsalgD as a 2.7-kb PCR product in pCR2.1 | This study |
| pAPDP | Cmr; 7.4-kb contains PsalgD as a 2.7-kb HindIII-EcoRV fragment in pBBR.Gus | This study |
| pAPDPΔ15 | Cmr; derivative of pAPDP containing a 1.5-kb deletion from the 5′ end of PsalgD | This study |
| pAPDPΔ23 | Cmr; derivative of pAPDP containing a 2.3-kb deletion from the 5′ end of PsalgD | This study |
| pSK3 | Smr Spr; contains a 1.0-kb fragment from PsalgD in pRG960sd in the transcriptionally active orientation | 44 |
| pSK4 | Smr Spr; contains a 1.0-kb fragment from PsalgD in pRG960sd in the transcriptionally inactive orientation | 44 |
| pAP32 | Tcr Kmr; contains Tn5-inactivated alginate genes from FF5.32 in pRK7813 | This study |
| pAP32.1 | Apr Kmr; contains a 5.3-kb BamHI fragment consisting of 2.8 kb from Tn5 and 2.5 kb of FF5.32 DNA in pBluescript SK(+) | This study |
| pMF4 | Tcr; cosmid clone from FF5(pPSR12) in pRK7813 | This study |
| pMF6 | Tcr; cosmid clone from FF5(pPSR12) in pRK7813 | This study |
| pMF6.1 | Apr; contains a 2.7-kb EcoRI fragment from pMF6 | This study |
| pMF6.2 | Apr; contains a 2.0-kb PstI fragment from pMF6 | This study |
| pMF6.21 | Tcr; 2.0-kb PstI fragment from pMF6.2 in pRK415 in the transcriptionally active orientation with respect to lacZ and algR1 | This study |
| pMF6.22 | Tcr; 2.0-kb PstI fragment from pMF6.2 in pRK415 in the transcriptionally inactive orientation with respect to lacZ and algR1 | This study |
| pAD1039 | Tcr; contains algR1 from P. aeruginosa in pCP13 | V. Kapatral |