Table 1.
Bacterial strains and plasmids
| Strain or plasmid | Genotype and/or phenotype | Source or reference |
|---|---|---|
| E. coli | ||
| DH5α | φ80dlacZΔM15Δ (lacZYA–argF) endA1 recA1 | Stratagene |
| P. aeruginosa | ||
| FRD1 | Cystic fibrosis isolate, Alg+ | Ohman & Chakrabarty (1981) |
| FRD2 | algT18, Alg− | DeVries & Ohman (1994) |
| FRD1108 | alg8Δ:: GmR (non-polar), Alg− | This study |
| FRD1144 | alg44:: GmR (non-polar), Alg− | This study |
| FRD1200 | algG:: GmR (non-polar), Alg− | Jain et al. (2003) |
| PAO1 | Wild-type, non-mucoid | This laboratory |
| Plasmids | ||
| pEX100T | oriV(ColE1) bla (CbR/ApR) sacB oriT | Schweizer (1992) |
| pPHO7 | P15A oriV (TcR) ‘phoA (2.6 kb XbaI–XhoI fragment) | Gutierrez & Devedjian (1989) |
| pMF54 | ColE1-SF broad-host replicon ApR oriT(RK2) Ptrc lacIQ | Franklin et al. (1994) |
| pRK2013 | ColE1 Tra(RK2)+ KmR | Jain & Ohman (1998) |
| pSJ12 | Source of GmR non-polar cassette (0.7 kb) | Jain & Ohman (1998) |
| pSJ44 | pMF54 expressing alg44 algK (3.8 kb EcoRI fragment) | This study |
| pSJ184 | pEX100T with Δalg8:: GmR alg44 | This study |
| pSJ199 | pMF54 expressing alg8 alg44 (3.6 kb XhoI–KpnI fragment) | This study |
| pSJ220 | pEX100T with alg44:: GmR algK | This study |
| pSJ340 | pMF54 expressing alg8(1-346)-phoA | This study |
| pLO11 | pMF54 expressing alg8(1-417)-phoA | This study |
| pLO12 | pMF54 expressing alg8(1-393)-phoA | This study |
| pLO17 | pMF54 expressing alg8(1-377)-phoA | This study |
| pLO19 | pMF54 expressing alg8(1-34)-phoA | This study |
| pLO22 | pMF54 expressing alg8(1-245)-phoA | This study |
| pLO25 | pMF54 expressing alg8(1-474)-phoA | This study |
| pLO28 | pMF54 expressing alg8(1-494)-phoA | This study |
| pLO37 | pMF54 expressing alg8(D136A) | This study |
| pLO39 | pMF54 expressing alg8(D191A) | This study |
| pLO42 | pMF54 expressing alg8(D299A) | This study |
| pLO44 | pMF54 expressing alg8(L339A) | This study |
| pLO63 | pMF54 expressing alg8(V158A) | This study |
| pLO71 | pMF54 expressing alg8(R342A) | This study |
| pLO77 | pMF54 expressing alg8(W343A) | This study |
| pLO32 | pMF54 expressing alg44(1-35)-phoA | This study |
| pLO33 | pMF54 expressing alg44(1-139)-phoA | This study |
| pLO34 | pMF54 expressing alg44(1-250)-phoA | This study |
| pLO51 | pMF54 expressing alg44(1-389)-phoA | This study |
| pLO66 | pMF54 expressing alg44(1-364-stop) | This study |
| pSJ295 | pMF54 expressing alg44(1-244)-phoA | This study |
| pSJ303 | pMF54 expressing alg44(1-142)-phoA | This study |
Abbreviations: Alg+, mucoid; Alg−, non-mucoid; ApR, ampicillin resistance; CbR, carbenicillin resistance; GmR, gentamicin resistance (non-polar); KmR, kanamycin resistance; TcR, tetracycline resistance. phoA, promoterless alkaline phosphatase gene.