TABLE 1.

Bacterial strains, plasmids, and primers used in this study

Strain, plasmid, or primerGenotype, sequence, or propertiesReference
S. maltophilia
    KJWild type; a clinical isolate from Taiwan 9
    KJΔDI S. maltophilia KJ ampDI isogenic mutant; deletion of 103-bp internal DNA fragment of ampDI geneThis study
    KJΔDII S. maltophilia KJ ampDII isogenic mutant; deletion of 149-bp internal DNA fragment of ampDII geneThis 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, respectivelyThis study
    KJΔR S. maltophilia KJ ampR isogenic mutant; deletion of 468-bp internal DNA fragment of ampR geneThis study
E. coli
    DH5αλ ϕ80dlacZ&Dgr;M15 &Dgr;(lacZYA-argF)U169 recA1 endA1 hsdR17(rK mK) supE44 thi-1 gyrA relA1Invitrogen
    S17-1λ pir+ mating strain
Plasmids
    pEX18Tc sacB oriT Tcr 6
    pAmpDIpEX18Tc vector with an 820-bp DNA fragment of S. maltophilia KJ containing the intact ampDI gene and the 50-bp upstream region; TcrThis study
    pAmpDIIpEX18Tc vector with a 2,172-bp DNA fragment of S. maltophilia KJ containing the intact ampDII gene and the 858-bp upstream region; TcrThis study
    pAmpDIIDIpEX18Tc vector with a 2,502-bp DNA fragment containing the intact ampDI and ampDII genes; TcrThis study
    pΔDIpEX18Tc 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; TcrThis study
    pΔDIIpEX18Tc 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; TcrThis study
Primersa
    AmpDI-F5′-CGAAGCTTCGACAAGGAAAGGGAAGGCAG-3′This study
    AmpDI-R5′-CAAGATCTGCACCCACCAACAGCGGCAG-3′This study
    AmpDII-F5′-CACTTCCACTGTCCTCGTTC-3′This study
    AmpDII-R5′-CCCTTGCCCTTCAGTTCC-3This study
    DIN-F5′-TGAAGCTTCCAATGGTGGCAGTGG-3′This study
    DIN-R5′-GGTCTAGAAGTGGCAGGCGGTCTTC-3′This study
    DIC-F5′-GGTCTAGACAACAGCGGGCATTTCTAC-3′This study
    DIC-R5′-CTGAATTCCGCACGCATCTACGCCGAC-3′This study
    16rDNAQ-F5′-GACCTTGCGCGATTGAATG-3′This study
    16rDNAQ-R5′-CGGATCGTCGCCTTGGT-3′This study
    L1Q-F5′-ACCCCTGGCAGATCGGCAC-3′This study
    L1Q-R5′-CAGCAGCACCGCCGTTTC-3′This study
    L2Q-F5′-AACGCACCCACCGATGCC-3′This study
    L2Q-R5′-CGCCTGTCCAGCAATGCC-3′This study
    AmpDIQ-F5′-CTACGAAGACCGCCTGCC-3′This study
    AmpDIQ-R5′-GAAATGCCCGCTGTTGCC-3′This study
    AmpDIIQ-F5′-CCACCACACCGAGCAGAAG-3′This study
    AmpDIIQ-R5′-ATCTGCGCCGCACTGAAC-3′This study
  • a Underlining indicates the restriction sites introduced for cloning.