TABLE 1.
Bacterial strains, plasmids, and primers used in this study
| Strain, plasmid, or primer | Genotype, sequence, or properties | Reference |
|---|---|---|
| S. maltophilia | ||
| KJ | Wild type; a clinical isolate from Taiwan | 9 |
| KJΔDI | S. maltophilia KJ ampDI isogenic mutant; deletion of 103-bp internal DNA fragment of ampDI gene | This study |
| KJΔDII | S. maltophilia KJ ampDII isogenic mutant; deletion of 149-bp internal DNA fragment of ampDII gene | This study |
| KJΔDIΔDII | S. maltophilia KJ double mutant of ampDI and ampDII genes; deletion of 103-bp and 149-bp internal DNA fragments of ampDI and ampDII genes, respectively | This study |
| KJΔR | S. maltophilia KJ ampR isogenic mutant; deletion of 468-bp internal DNA fragment of ampR gene | This study |
| E. coli | ||
| DH5α | λ− ϕ80dlacZ&Dgr;M15 &Dgr;(lacZYA-argF)U169 recA1 endA1 hsdR17(rK− mK−) supE44 thi-1 gyrA relA1 | Invitrogen |
| S17-1 | λ pir+ mating strain | |
| Plasmids | ||
| pEX18Tc | sacB oriT Tcr | 6 |
| pAmpDI | pEX18Tc vector with an 820-bp DNA fragment of S. maltophilia KJ containing the intact ampDI gene and the 50-bp upstream region; Tcr | This study |
| pAmpDII | pEX18Tc vector with a 2,172-bp DNA fragment of S. maltophilia KJ containing the intact ampDII gene and the 858-bp upstream region; Tcr | This study |
| pAmpDIIDI | pEX18Tc vector with a 2,502-bp DNA fragment containing the intact ampDI and ampDII genes; Tcr | This study |
| pΔDI | pEX18Tc vector with a 1,918-bp DNA fragment of S. maltophilia KJ containing the ampDI gene with an internal 103-bp deletion, the 841-bp upstream region, and 349-bp downstream region of ampDI; Tcr | This study |
| pΔDII | pEX18Tc vector with a 2,089-bp DNA fragment of S. maltophilia KJ containing the ampDII gene with an internal 149-bp deletion, the 889-bp upstream region, and 523-bp downstream region of ampDI; Tcr | This study |
| Primersa | ||
| AmpDI-F | 5′-CGAAGCTTCGACAAGGAAAGGGAAGGCAG-3′ | This study |
| AmpDI-R | 5′-CAAGATCTGCACCCACCAACAGCGGCAG-3′ | This study |
| AmpDII-F | 5′-CACTTCCACTGTCCTCGTTC-3′ | This study |
| AmpDII-R | 5′-CCCTTGCCCTTCAGTTCC-3 | This study |
| DIN-F | 5′-TGAAGCTTCCAATGGTGGCAGTGG-3′ | This study |
| DIN-R | 5′-GGTCTAGAAGTGGCAGGCGGTCTTC-3′ | This study |
| DIC-F | 5′-GGTCTAGACAACAGCGGGCATTTCTAC-3′ | This study |
| DIC-R | 5′-CTGAATTCCGCACGCATCTACGCCGAC-3′ | This study |
| 16rDNAQ-F | 5′-GACCTTGCGCGATTGAATG-3′ | This study |
| 16rDNAQ-R | 5′-CGGATCGTCGCCTTGGT-3′ | This study |
| L1Q-F | 5′-ACCCCTGGCAGATCGGCAC-3′ | This study |
| L1Q-R | 5′-CAGCAGCACCGCCGTTTC-3′ | This study |
| L2Q-F | 5′-AACGCACCCACCGATGCC-3′ | This study |
| L2Q-R | 5′-CGCCTGTCCAGCAATGCC-3′ | This study |
| AmpDIQ-F | 5′-CTACGAAGACCGCCTGCC-3′ | This study |
| AmpDIQ-R | 5′-GAAATGCCCGCTGTTGCC-3′ | This study |
| AmpDIIQ-F | 5′-CCACCACACCGAGCAGAAG-3′ | This study |
| AmpDIIQ-R | 5′-ATCTGCGCCGCACTGAAC-3′ | This study |
a
Underlining indicates the restriction sites introduced for cloning.