TABLE 1.

Primers used for PCR amplification of the ftsI gene and site-directed mutagenesis

UseForward primeraReverse primera
PCR amplification of complete ftsI gene5′-CTCGTTATCCGTTACAGCAG-3′5′-GCCAAACCGTGTGATGAAAC-3′
Introduction of R517H change5′-AAAACGGGTACGGCACATAAGATTGAA-3′5′-TTCAATCTTATGTGTGCCGTACCCGTTTT-3′
Introduction of N526K change5′-GGACATTATGTAAAGAAGTATGTGGCA-3′5′-TGCCACATACTTCTTTACATAATGTCC-3′
Introduction of M3771 change5′-GAGATTTTAATTAACTCAAGTAACCGTGGT-3′5′-ACCACGGTTACTTGAGTTAATTAAAATCTC-3′
Introduction of S385T change5′-AACCGTGGTGTAACTCGTCTTGCATTA-3′5′-TAATGCAAGACGAGTTACACCACGGTT-3′
Introduction of S385T + L389F changes5′-GGTGTAACTCGTCTTGCATTTCGTATGCCA-3′5′-TGGCATACGAAATGCAAGACGAGTTACACC-3′
  • a Oligonucleotide sequences were based on the H. influenzae Rd genomic sequence (GenBank accession number NC000907).