TABLE 3.
Primer pairs used for gene inactivationa
| Gene | Primer name | Sequence (5′–3′) |
|---|---|---|
| rfmA | rfmA-UF | TAGATATCGCTCGGCACAGACGGAGAAC |
| rfmA-UR | atccccgggtaccgaGGCGTAGTAGGCGGTGAG | |
| rfmA-Km-F | cgcctactacgccTCGGTACCCGGGGATCCG | |
| rfmA-Km-R | gccaggtgggagtTGCAGGTCGACTCTAGAGATGC | |
| rfmA-DF | gagtcgacctgcaACTCCCACCTGGCGTCCTC | |
| rfmA-DR | TAGAATTCCGCCCAGTGTCTCCAGTTC | |
| rfmA-CF | ACGAACGGACCACCGACACC | |
| rfmA-CR | GCCGAGGAGAACACGACGAA | |
| rfmB | rfmB-UF | TAGATATCCCAGCAGTCCGTCATCAACCA |
| rfmB-UR | ccccgggtaccgaATCGAGTGACCGGCCAGCAC | |
| rfmB-Km-F | cggtcactcgatTCGGTACCCGGGGATCCG | |
| rfmB-Km-R | gccgggatacctTGCAGGTCGACTCTAGAGATGC | |
| rfmB-DF | gagtcgacctgcaAGGTATCCCGGCTCAAGGTCAG | |
| rfmB-DR | TAGAATTCACAGCGGTGCGTCCATGTCG | |
| rfmB-CF | TGCATCTGGCGATGGTGGC | |
| rfmB-CR | CGACTGCTCGAATCCGGTGA | |
| rfmD | rfmD-UF | TAGATATCCACCTCGTGTCGCACCCT |
| rfmD-UR | ccccgggtaccgaAGTTCATCCGCCAGCTCAC | |
| rfmD-Km-F | ggcggatgaactTCGGTACCCGGGGATCCG | |
| rfmD-Km-R | aggtgacgcggtTGCAGGTCGACTCTAGAGATGC | |
| rfmD-DF | gagtcgacctgcaACCGCGTCACCTGGCAAC | |
| rfmD-DR | TAGAATTCGCCGGACACGGAGGAGAAC | |
| rfmD-CF | CGGTTCGCCGTGTCGTCCTT | |
| rfmD-CR | CGGGTTCTGTGCGGTTGTGC | |
| rfmE | rfmE-UF | TAGATATCTGCGGGAGATGCTGTTCGG |
| rfmE-UR | ccccgggtaccgaGTGTCACCAAGGCTGGAGTGGA | |
| rfmE-Km-F | ccttggtgacacTCGGTACCCGGGGATCCG | |
| rfmE-Km-R | tcgtcgccgccgTGCAGGTCGACTCTAGAGATGC | |
| rfmE-DF | gagtcgacctgcaCGGCGGCGACGAGATGTT | |
| rfmE-DR | TAGAATTCGCCTCAAGGGCCTCACGATC | |
| rfmE-CF | GGCTGGCTTTCCTGTTCTCGG | |
| rfmE-CR | CGGTCACTGTGGTGGTCTTGC | |
| rfmF | rfmF-UF | TAGATATCGTGGCTCGGCTCGGTGAAGT |
| rfmF-UR | ccccgggtaccgaTGGCTGACCCGCAGATAGCG | |
| rfmF-Km-F | gcgggtcagccaTCGGTACCCGGGGATCCG | |
| rfmF-Km-R | tgggtcacgcccTGCAGGTCGACTCTAGAGATGC | |
| rfmF-DF | gagtcgacctgcaGGGCGTGACCCACTTCCTG | |
| rfmF-DR | TAGAATTCGGCCTGATCCGCGTCCAGT | |
| rfmF-CF | CCGCGTGTCGTACACCTTCG | |
| rfmF-CR | GAACACCGCCTCCCAGTCCA |
a
The underlined letters represent the restriction sites of EcoRV and EcoRI. The letters in lowercase are reverse complement sequences for overlap extension PCR.