Table 2 Primers used for PCR amplification, site directed mutagenesis and sequencing of the ftsI and acrR genes
From: A novel PBP3 substitution in Haemophilus influenzae confers reduced aminopenicillin susceptibility
Use of primers | Forward primer sequence | Reverse primer sequence |
|---|---|---|
Ampification of full length ftsI gene | 5′ – CCTGCGTGTTTGAAAGTTGAAAGAGATG – 3’ | 5′ – AACAAAGTAAGGGCGAGGATATTCCCAAAG – 3’ |
Introduction of Y528H substitutiona in ftsI in NTHi3655 | 5′ – GAAAATGGACATTATGTAAATAAGCATGTGGCATTTACTGCGGG – 3’ | 5′ – CCCGCAGTAAATGCCACATGCTTATTTACATAATGTCCATTTTC – 3’ |
Amplification and sequencing of the acrR gene | 5′ – TTGTGGGTTTACGGCTTACC – 3’ | 5′ – CCGATGACACCGACAAAAAT – 3’ |
Sequencing of ftsI gene fw1 (using primer walking) | 5′ – CCAATAAACTCTACAGTTAAATGCTCGC – 3’ | |
Sequencing of ftsI gene fw2 (using primer walking) | 5′ – AGCGGACGATAAACACCGAAACTACCA – 3’ | |
Sequencing of ftsI gene fw3 (using primer walking) | 5′ – ATACTTAAGGTAACATCTTGTGCATCATAT – 3’ |