Table 2.

Oligonucleotide primers used in this study

PrimerSequenceaTarget site
MiaBP15′-ATAGCCAAGCTT CGGTAGCTTAAGTTC-3′ E. coli o391 (nt 8697–8671 in accession no. AE000170)
MiaBP25′-TTCCAG GGATCCAACTGATGGAAGAC-3′ E. coli miaB (nt 8032–8057 in AE000170)
MiaBP35′-GGTGTAAGGATCCACGCAGTAGGTGC-3′ E. coli miaB (nt 7725–7750 in AE000170)
miaBPstI5′-AGGCCCACCGGAACTGCAGGCCTG-3′4 bp downstream of the UAA codon of E. coli miaB (nt 6779–6802 in AE000170)
miaBSmaI5′-ATTTTTACCCC*GGGCTGATAACGCTCA-3′105 bp upstream of E. coli miaB AUG start codon (nt 8336–8307 in AE000170)
MiaBT15′-AAGGTGGATCC GTTCCGGTTCGATATC-3′ E. coli f359a (nt 6394–6420 in AE000170)
MiaBT25′-ATCATG AAGCTTTCCGGGCGTACGG-3′ E. coli miaB (nt 7043–7019 in AE000170)
MiaBT35′-TCCGTAAGCTT CGTGCGGCGCGTCC-3′ E. coli miaB (nt 7363–7339 in AE000170)
MotH5′-AGGGACGGCTGACTTCTTCACC-3′ S. typhimurium miaB(nt 1165–1144 in AJ249116)
PCR15′-AATACCGAGACGACGTTCGAGCTG-3′ S. typhimurium f359a(nt 2376–2353 in AJ249116)
PCR25′-AAATCCGCACGTCAGGCTGGCTG-3′ S. typhimurium o391(nt 334–356 in AJ249116)
Sekv145′-TACTCGTTCATCTGACAGCC-3′ S. typhimurium miaB(nt 683–664 in AJ249116)
T-L5′-ACCTTTGGTCACCAACGCTTTTCC-3′ Tn10dTc, ∼70 bp from the left end, facing outward
Tn10XbaI5′-TAATCTAGAACCTCTTACGTGCC-3′ Tn10dCm, 342 bp from the end, facing outward
  • a The underlined bases represent synthetic restriction sites. Base changes introduced to create the restriction sites are in boldface. The ∗ in miaBSmaI means that there are three additional bases (TAA) in this place in the E. coli chromosomal sequence.