TABLE 1.

Primers used in this study

PrimerSequence 5′→3′Amplification target
Northern blot probe PCR
    Sta2 ForGCGTATGTTCAATTTGTCG1,140-bp product from lst of N. gonorrhoeae and N. meningitidis
    Sta3 RevCGTCAAATGTCAAAATCGG
Real-time PCR
    LstRTFAAACCCGCATACGAGGTATGA100-bp product from lst of N. gonorrhoeae and N. meningitidis
    LstRTRAAGCCGGTTTCAATGCGTAA
    16S RT FGCGTGGGTAGCAAACAGGAT100-bp product from rrs gene of N. gonorrhoeae and N. meningitidis
    16S RT RCGCGTTAGCTACGCTACCAAG
Cloninga
    FusBam F1CGCTGGATCCGACATCAATATCGG496 bp of N. gonorrhoeae 5′lst including RBS sequence and 24 bp of lst including ATG
    FusBam R1CAAAGGATCCTTTTTCAAGCCC586 bp of N. meningitidis 5′lst including RBS sequence and 24 bp of lst including ATG
Primer extension
    PE1CGCATTCCTTTCCCCCTGATTTAC3′ region of primers anneals 78, 16, and 24 bp downstream of lst ATG, respectively
    PE2CACAACACGGTCAAACAAGC
    PE3CAATCAGGCACAACACGGTC
RPA primers
    PA1GATCGAGCTCGTTCGATCTTGGCGTGTTTG292 bp of N. gonorrhoeae 5′lst excluding the RBS sequence; SacI site of PA1 is denoted by double underline
    PA2GATCGGATCCCTCCATTCCGACAAATTGAAC
    PA3AATTAACCCTCACTAAAGGG397-bp product from pSKII including the cloned 292-bp N. gonorrhoeae 5′lst insert
    PA4GTAATACGACTCACTATAGGG
RT-PCR: Fig. 7
    P1CGGGATCCGGCTTTCCCGCGTTTGCCGG5′ region of the primer anneals upstream of CREE; 282 bp upstream of lst ATG
    P2CGGGATCCCGCCTTGTGCCTGATGTGCG5′ region of the primer anneals upstream of CREE; 252 bp upstream of lst ATG
    P3CGGGATCCTTTCGGTAAAATTGATTTTA5′ region of the primer anneals upstream of CREE; 222 bp upstream of lst ATG
    P4CGGGATCCAACTGTCGGAATATCTGCTA5′ region of the primer anneals downstream of CREE; 91 bp upstream of lst ATG
    P5CGGGATCCTTTTTCCGTCCCGGGACAC5′ region of the primer anneals downstream of CREE; 61 bp upstream of lst ATG
    P6CGGGATCCACACTCGGGGCGTATGTTCA5′ region of the primer anneals downstream of CREE; 41 bp upstream of lst ATG
    P7CGGGATCCAGGGATATGGGCTTGAAAAAG5′ region of the primer anneals downstream of CREE; 6 bp upstream of lst ATG
    CrevCTCCGCCATCGTCGGAATCommon reverse primer used in conjunction with primers P1 to P7 and the 3′ region of the primer anneals 372 bp downstream of lst ATG
RT-PCR: Fig. 8
    IP1TTATTCTCTCTTGTAGGTTGGGenerates a 212-bp product of 5′icd upstream region; P4 and Crev primers used in Fig. 7 were used in Fig. 8
    IP2TGCCGCCGCACATCAGGCACA
  • a Primers FusBam F1 and FusBam R1 were used to include regions of the N. gonorrhoeae or N. meningitidis 5′lst region including the RBS and 24 bp of the lst gene including ATG to create translational lacZ fusions in pLES94. The BamHI restriction sites are underlined with single lines.