TABLE 2.

Oligonucleotide primers

Primer function and nameSequence (5′-3′)UsageGenBank accession no. (nt) or source
Cloning and real-time PCR
    Int-FCGTTCCATACAGAAGCTGCloning of the In58-G1 integron EU598463 (1-18)
    3-CSAAGCAGACTTGACCTGACloning of the In58-G1 integron EU598463 (1625-1641)
    Ven-8PGTAATCTCTCTCCTGGGCTRT-PCR for blaGES-1 EU598463 (1343-1361)
    Ven-12CCCCAAGGAGAGATCGTRT-PCR for blaGES-1 EU598463 (744-762)
    Aph-FGGAAACGTCTTGCTCGAGGRT-PCR for aphA1 X06402 (1940-1958)
    Aph-RACTGAATCCGGTGAGAATGGRT-PCR for aphA1 X06402 (2480-2500)
Site-directed mutagenesis
    P1st-FaAACCCAGTTGACATAAGCCTGTTCGGTTCGTAAACTGTAATGConstruction of the strong promoter from the weak promoterThis study
    P1st-RaCATTACAGTTTACGAACCGAACAGGCTTATGTCAACTGGGTTConstruction of the strong promoter from the weak promoterThis study
    P1h1-FbGCACGAACCCAGTGGACATAAGCCTGTTCGConstruction of hybrid 1 from the weak promoter (−35 box)This study
    P1h1-RbCGAACAGGCTTATGTCCACTGGGTTCGTGCConstruction of hybrid 1 from the strong promoter (−35 box)This study
    P1h2-FbGCACGAACCCAGTTGACATAAGCCTGTTCGConstruction of hybrid 2 from the weak promoter (−35 box)This study
    P1h2-RbCGAACAGGCTTATGTCAACTGGGTTCGTGCConstruction of hybrid 2 from the weak promoter (−35 box)This study
    P2ac-FcTGACTGTTTTTTTGGGGTACAGTCTATGCCTCGGGAConstruction of the P2 promoterThis study
    P2ac-RcTGCCCGAGGCATAGACTGTACCCCAAAAAAACAGTCAConstruction of the P2 promoterThis study
    PC-FdGGATGAAGGCACGAACCCAGAGCTTACGAACCGAACAGTGTCCAGTAATGCAAGTAGCGTATGCConstruction of a promoterless sequence upstream of the blaGES-1cassetteThis study
    PC-RdGCATACGCTACTTGCATTACTGGACACTGTTCGGTTCGTAAGCTCTGGGTTCGTGCCTTCATCCConstruction of a promoterless sequence upstream of the blaGES-1cassetteThis study
  • a Underlined bases correspond to the −35 and −10 hexamers of the Pc strong promoter.

  • b Underlined bases correspond to the −35 hexamer of the hybrid Pc promoters.

  • c Underlined bases correspond to the insertion of three G bases in the P2 promoter sequence.

  • d Bases in boldface type correspond to the reversed Pc weak promoter sequence. The −10 and −35 hexamers are underlined.