Table 1.

Oligonucleotide primers used to amplify the type II topoisomerase genes from S. pneumoniae for protein purification and DNA sequencing

ApplicationGeneForward primer (nucleotide positions)Reverse primer (nucleotide positions)
Protein purification gyrA 5′-ATGCAGGATAAAAATTTACTG (bp 1–21)5′-CTGATTTGTATTAATAAAATTGTTTAT (158–185 bp downstream of stop codon)
gyrB 5′-ATGACAGAAGAAATCAAAAATCTG (bp 1–24)5′-CATATTTTCTAGACCAAGGGAAC (17–39 bp downstream of stop codon)
parC 5′-ATGTCTAACATTCAAAACATGTC (bp 1–23)5′-TTATTTATCTTCAGTAACTACTTC (7–30 bp downstream of stop codon)
parE 5′-ATGTCAAAAAAAGGAAATCAAT (bp 1–23)5′-TTAAAACACTGTCGCTTCTTC (1–22 bp downstream of stop codon)
Target gene amplification gyrA 5′-TAAAAAACTTTGTCACGAATATGCC (130–105 bp upstream of start)5′-AACGATACGCTCACGACCAGT (bp 750–771)
gyrB 5′-TGAAGGACAAACCAAGACCAAA (bp 1032–1053)5′-GTCCATTTCACCTAGCCCCTTATA (bp 1759–1782)
parC 5′-AAACCTACTCTACATTCTTTGAAAGGAG (134–106 bp upstream of start)5′-CAGTTGGGTGGTCAATCATGTAAA (bp 571–594)
parE 5′-ACCAAGGATAAACATGGAAGCC (bp 1026–1047)5′-CATTCATCTCACCAAGTCCTTTGTA (bp 1737–1762)
QRDR amplification gyrA 5′-CGTTTTAGTGGTTTAGAGGC (85–66 bp upstream of start)5′-GACCAACTTCACTGCATCA (bp 567–585)
gyrB 5′-TTCTCCGATTTCCTCATG (bp 1105–1122)5′-CCCGGCTGGATATATTCT (bp 1665–1682)
parC 5′-CGCCCTAGATACTGTGTGA (98–80 bp upstream of start)5′-AAATCCCAGTCGAACCAT (bp 493–510)
parE 5′-TGTGGATGGAATAGTGGC (bp 1054–1071)5′-ACCGAACTGTTTACGGAGT (bp 1697–1715)