Table 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Descriptiona | Source or reference |
|---|---|---|
| Strains | ||
| B. multivorans strains | ||
| ATCC 17616 | Type strain | ATCC |
| 26D7 | Polymyxin B-susceptible derivative of ATCC 17616 with Tn5-751S disrupting Bmul_2133; TMPr | This study |
| RMI17 | ΔBmul_2134 derivative of ATCC 17616; TETs | This study |
| RMI18 | ΔBmul_2134 derivative of ATCC 17616; TETs | This study |
| RMI19 | ΔBmul_2134 derivative of ATCC 17616; TETr | This study |
| P. aeruginosa strains | ||
| ATCC 27853 | MIC quality control strain | ATCC |
| M2 | Mouse intestinal isolate susceptible to neutrophil killing | 44 |
| E. coli strains | ||
| DH5α | F− ϕ80dlacZΔM15 Δ(lacZYA-argF)U169 deoR recA1 endA1 hsdR17(rK− mK−) phoA supE44 λ−thi-1 gyrA96 relA1 | Invitrogen |
| HB101 | supE44 hsdS20(rB mB) recA13 ara-14 proA2 lacY1 galK2 rpsL20 xyl-5 mtl-1 | 40 |
| XL1-Blue MR | Cosmid library host strain | Agilent Technologies |
| Plasmids | ||
| pBluescript II SK(+) | Cloning vector; AMCr | Agilent Technologies |
| Super Cos 1 | Cosmid cloning vector; AMCr NEMr | Agilent Technologies |
| pCR-BluntII-TOPO | Cloning vector; KANr ZEOr | Invitrogen |
| pTGL166 | Broad-host-range temperature-sensitive IncP1 plasmid containing Tn5-751S; KANr TMPr TETr PENr | 7 |
| pRK2013 | ColE1 Tra (RK2)+ KANr | 14 |
| pEX18Tc | Suicide vector; TETr | 18 |
| pUCP26 | Broad-host-range vector; TETr | 47 |
| pUCP28T | Broad-host-range vector; TMPr | 42 |
| pBS1 | pBluescript II KS(+) containing Tn5-751 and flanking DNA from mutant 26D7; AMPr KANr | This study |
| pBS4 | pBluescript II KS(+) carrying a 4.0-kb BamHI fragment containing Bmul_2134; AMCr | This study |
| pRM301 | pEX18Tc carrying the 4.0-kb BamHI fragment from pBS4 containing Bmul_2134; TETr | This study |
| pRM302 | pRM301 with a 718-bp NotI fragment deletion in Bmul_2134; TETr | This study |
| pRM305 | pUCP28T with a 1.6-kb AfeI/ApaI fragment from pBS4 blunt ligated into SmaI with Bmul_2134; TMPr | This study |
| pRM306 | pCR-BluntII-TOPO with a 1.6-kb Bmul_2133 PCR fragment; TETr | This study |
| pRM308 | pUCP26 with a 1.6-kb XbaI/HindIII fragment from pRM306 containing Bmul_2133; TETr | This study |
a
AMC, ampicillin; KAN, kanamycin; NEM, neomycin; PEN, penicillin; TET, tetracycline; TMP, trimethoprim; ZEO, zeocin.