Table 2.

PCR primers used in sequence determination and mutant construction

PrimerNucleotide sequenceaAmplicon size (bp)
ComC-F55′-AGTTTTTTGTCTGGCTGCG-3′651
ComC-B55′-TCCACTAAAGGCTCCAATCG-3′
ComC′-F15′-GACTAGTCATTGGCGGAAGCGGAAGCCTATCAAC-3′3,423
ComC′-B15′-GCTCTAGAGCTCAGAACATCAAAAATGACCGTTTAGGAC-3′
Erm2-F35′-GACTAGTCCAAACAGGTAACGGTTATTGCAGG-3′1,291
Erm2-B15′-GCTCTAGAGCCCTCTTTAGCTCCTTGGAAGCTGT-3′
ComD-F15′-CGGGATCC CGCTAAGTTACCTCTTTTCTCAGTG-3′292
ComD-B15′-GGAATTCC GCTTCCTTTTGTGCCATTATC-3′
ComE-F15′-CGGGATCC CCTGAAAAGGGCAATCACCAG-3′462
ComE-B15′-GGAATTCCGCGATGGCACTGAAAAAGTCTC-3′
  • a Engineered restriction sites are underlined; actual endonuclease recognition sequences are in boldface. Restriction endonuclease recognition sequences are as follows:BamHI, G/GATCC; EcoRI, G/AATTC; Spel, A/CTAGT; Xbal, T/CTAGA.