TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Relevant characteristic(s)a | Reference or source |
|---|---|---|
| Escherichia coli | ||
| DH5α | supE44 ΔlacU169 (φ80 lacZΔM15) hsdR17 recA1 endA1 gyrA96 thi-1 relA1 | 59 |
| HB101 | supE44 hsdS20(rB mB) recA13 ara-14proA2 lacY1 galK2 rpsL20 xyl-5 mtl-1 | 5 |
| Pseudomonas syringae pv. glycinea | ||
| PG4180 | Wild type, produces small amounts of alginate | R. Mitchell |
| PG4180.muc | Emerged spontaneously from PG4180, produces vast amounts of alginate | This study |
| Azotobacter vinelandii | ||
| wtC15 | Produces alginate | Tonje Strommen, NTNU,b Trondheim, Norway |
| Pseudomonas aeruginosa | ||
| Pa8821 | Alg+ (unstably mucoid) | 12 |
| Pa8822 | Alg− (nonmucoid revertant of Pa8821) | 12 |
| Pa8830 | Alg+ (stably mucoid) | 12 |
| Plasmids | ||
| pBluescript SK(+) | Apr | Stratagene |
| pBlueSK:AXSalgTop | Apr; contains an 8,860-bp SalI fragment carrying the algT-mucABD gene cluster region from PG4180 cloned in pBluescript SK(+) | This study |
| pRK2013 | Kmr; helper plasmid | 20 |
| pBBR1MCS-3 | Tcr; ColE1 origin | 36 |
| pBBR3-AXSalgTop | Tcr; contains a 5,096-bp XbaI-HindIII PCR fragment (consisting of 988-bp upstream algT promoter region, 582-bp algT reading frame, 591-bp mucA reading frame, 960-bp mucB reading frame, and 1,440-bp mucD reading frame) from PG4180.muc cloned in pBBR1MCS-3 | This study |
| pBBR3-AXSalgT | Tcr; contains a 1,837-bp XbaI-PmlI PCR fragment (consisting of 988-bp upstream algT promoter region, 582-bp algT reading frame, and 270-bp downstream algT region) from PG4180.muc cloned in pBBR1MCS-3 | This study |
a
Ap, ampicillin; Km, kanamycin; Tc, tetracycline.
b
NTNU, Norwegian University of Science and Technology.