Table 1.
Strain/plasmid | Genotype | Reference |
---|---|---|
E. coli | ||
DH5α | F−φ80dlacZΔM15 Δ(lacZYA–argF) U169 deoR recA1 endA1 hsdR17(rk−, mk+) phoA supE44 λ- thi-1 gyrA96 relA1 | New England Biolabs |
TOP10F′ | F′[lacIq, Tn10(TetR)] mcrA Δ(mrr-hsdRMS-mcrBC)φ80dlacZΔM15 ΔlacX74 deoR recA1 araD139 D(ara-leu)7697 galU galK rpsL (StrR) endA1 nupG | Invitrogen |
P. aeruginosa | ||
PAO1 | Prototype; Alg− | Holloway & Morgan (1986) |
PDO300 | PAOmucA22; Alg+ | Mathee et al. (1999) |
PKM805 | PAOmucA22algT24-1; Alg− | PDOalgT; PDO300 derivative; Ramos et al. (2003) |
PKM300 | PAOampR : : aacCI; Alg− | PAOampR; Kong et al. (2005b) |
PKM307 | PAOmucA22 ampR : : aacCI; Alg+ | PDOampR; this study |
PKM308 | PAOmucA22 attB : : PampC–lacZ; TcR, Alg+ | PDO300 derivative; this study |
PKM309 | PAOmucA22 attB : : PampR–lacZ; TcR, Alg+ | PDO300 derivative; this study |
PKM310 | PAOmucA22 ampR : : aacCI attB : : PampC-lacZ; GmR, TcR, Alg+ | PDOampR derivative; this study |
PKM311 | PAOmucA22 ampR : : aacCI attB : : PampR–lacZ; GmR, TcR, Alg+ | PDOampR derivative; this study |
Plasmids | ||
Mini-CTX-lacZ | TcR; integration-proficient vector for single-copy chromosomal lacZ fusion | Becher & Schweizer (2000) |
pEX100T | ApR; sacB oriT | Schweizer & Hoang (1995) |
pGEMEX-1 | ApR; ColE1ori lacZα | Promega |
pKMG37 | ApR; pQF50 containing PalgT-lacZ transcriptional fusion | Mathee et al. (1997) |
pLP170 | ApR; lacZ transcriptional fusion vector that contains an RNase III splice sequence positioned between the MCS and lacZ | Preston et al. (1997) |
pLPLA | ApR; pLP170 containing PlasA-lacZ transcriptional fusion | Preston et al. (1997) |
pLPLB | ApR; pLP170 containing PlasB-lacZ transcriptional fusion | Preston et al. (1997) |
pLPR1 | ApR; pLP170 containing PrhlI-lacZ transcriptional fusion | Van Delden & Iglewski (1998) |
pME6030 | TcR; oriVpVS1oriVp15AoriT | Heeb et al. (2000) |
pPCS223 | ApR; pLP170 containing PlasI-lacZ transcriptional fusion | Van Delden & Iglewski (1998) |
pPCS1001 | ApR; pLP170 containing PlasR-lacZ transcriptional fusion | Pesci et al. (1997) |
pPCS1002 | ApR; pLP170 containing PrhlR-lacZ transcriptional fusion | Pesci et al. (1997) |
pQF50 | ApR; broad-host-range vector with promoterless lacZ | Farinha & Kropinski (1990) |
pRK2013 | KmR; ColE1ori-Tra (RK2)+ | Figurski & Helinski (1979) |
pSJ01 | ApR; pGEMEX-1 with a 1220 bp EcoRI–BamHI flanked fragment containing ampR | Kong et al. (2005b) |
pSJ06 | ApR; pME6030 with a 1220 bp EcoRI–BamHI flanked fragment containing ampR (referred to as pAmpR) | Kong et al. (2005b) |
pSJ07 | ApR; pEX100T derivative with ampR : : aacCI | Kong et al. (2005b) |
pSJ09 | ApR, GmR; pGEMEX-1 with a 330 bp EcoRI–BamHI flanked fragment containing ampC-ampR intergenic region | Kong et al. (2005b) |
pSJ10 | TcR; CTX-lacZ fused with ampC promoter, PampC | Kong et al. (2005b) |
pSJ11 | TcR; CTX-lacZ fused with ampR promoter, PampR | Kong et al. (2005b) |
pUCGm | ApR, GmR; pUC19 derivative containing gentamicin cassette | Schweizer (1993) |
Primers | ||
SBJ01ampRFor* | 5′-GGAATTCTGGCGAACAGCAGTGTGGAAGCGG-3′ | |
SBJ02ampRRev* | 5′-CGGGATCCATTCCAATCACAACCCCAACGCC-3′ | |
SBJ03ampCRFor* | 5′-GGAATTCTGAGGCCGCGCGGCAGACGCTTGAACA-3′ | |
SBJ04ampCRRev* | 5′-CGGGATCCCATGAGGATTGGCGTCCTTTG-3′ | |
DBS_QRTAmpRF | 5′-CATTGGCCTTCATCACCGGTTGTA-3′ | |
DBS_QRTAmpRR | 5′-GGTTTCTCATGCAGCCCACGACAA-3′ |
*The italicized portion of the sequence indicates a restriction site in a PCR product prepared with the primer.