Skip to main content
. 2014 Jul;196(13):2472–2480. doi: 10.1128/JB.01512-14

TABLE 2.

Primers used in this study

Category and primer name Sequence (5′ to 3′)a
Construction of suicide plasmid for pflB gene
    EF delta pfl XbaI F GCGTCTAGACGCTTGCATCTAAAAGGTTATGTG
    EF delta pfl HindIII R CTCCAAGCTTGGAGGCTCAATTCCTTCTAATGG
    pHY300PLK tet P (+) SalI FCCGGTCGACCGGGCCATATTGTTGTATAAGTG
    pHY300PLK tet R GCTTCTAGAGATCTGCAGGTCG
Confirmation of pflB gene disruption
    EF delta pfl check F GGGCACACAAGAAGTTATTCAGTC
    EF delta pfl check R GGCATATTGAGTTTCTTCACTTGG
Construction of reporter assay and plasmids
    EF ldhL1 Pro XbaI F AAGTCTAGACTTTCATTCTCCTTGGATTATAAGC
    EF ldhL1 Pro SalI R ATGGTCGACCATGTGTACCATTCCTTCCTCTAC
    EF pdhAB Pro XbaI F TGCTCTAGAGCAATGAATGTTCGTGATAAAACC
    EF pdhAB Pro SalI R ATGGTCGACCATTTTGTCACACAATCCTCTCTATTACG
    EF pflB Pro XbaI F CCCTCTAGAGGGCACACAAGAAGTTATTCAGTC
    EF pflB Pro SalI R ATGGTCGACCATGAAGTGTTTGCCTCCTTAGTT
    EF adhE Pro F GGTTAAGCCGAATGAATTTTTTGAC
    EF adhE Pro SalI R ATGGTCGACCATGTTCATCCTCCTAAGAGTTTCATT
    beta-gal SalI F GACGTCGACGTCGTTTTACAACGTCGTGACTG
    beta-gal SD ATG-fuse XbaI F GTTTCTAGAAACTAAGGAGGCAAACACTTCATG
    beta-gal SphI R AGGGCATGCCCTGCCCGGTTATTATTATTTTTGA
RT-PCR
    EF RT-PCR 16S F GTGTCGTTGATGGATGGACC
    EF RT-PCR 16S R TCTACGCATTTCACCGCTACAC
    EF RT-PCR pflA F CCGTTTTATCGTATTTACACAAGGG
    EF RT-PCR pflA R GGCATCCAGACGGATTAAATATTC
    EF RT-PCR pflB F GAGAACAAGCGTTGACACTCG
    EF RT-PCR pflB R CCATATAATGCGATACGACGATAGTC
    EF RT-PCR glpF1 F CGAACAAAATACGCTGGTTCTG
    EF RT-PCR glpF1 R CAAACAAGCAAGCCAACAGC
    EF RT-PCR glpF2 F GGTTGGGTCGTTATTGCTTTAGG
    EF RT-PCR glpF2 R ATTGGCTGGATAGTTACGGACAG
a

Underlined nucleotides indicate restriction sites.