TABLE 2.
Oligonucleotide primers used in this study
| Primer | Sequencea | Target site |
|---|---|---|
| MiaBP1 | 5′-ATAGCCAAGCTTCGGTAGCTTAAGTTC-3′ | E. coli o391 (nt 8697–8671 in accession no. AE000170) |
| MiaBP2 | 5′-TTCCAGGGATCCAACTGATGGAAGAC-3′ | E. coli miaB (nt 8032–8057 in AE000170) |
| MiaBP3 | 5′-GGTGTAAGGATCCACGCAGTAGGTGC-3′ | E. coli miaB (nt 7725–7750 in AE000170) |
| miaBPstI | 5′-AGGCCCACCGGAACTGCAGGCCTG-3′ | 4 bp downstream of the UAA codon of E. coli miaB (nt 6779–6802 in AE000170) |
| miaBSmaI | 5′-ATTTTTACCCC*GGGCTGATAACGCTCA-3′ | 105 bp upstream of E. coli miaB AUG start codon (nt 8336–8307 in AE000170) |
| MiaBT1 | 5′-AAGGTGGATCCGTTCCGGTTCGATATC-3′ | E. coli f359a (nt 6394–6420 in AE000170) |
| MiaBT2 | 5′-ATCATGAAGCTTTCCGGGCGTACGG-3′ | E. coli miaB (nt 7043–7019 in AE000170) |
| MiaBT3 | 5′-TCCGTAAGCTTCGTGCGGCGCGTCC-3′ | E. coli miaB (nt 7363–7339 in AE000170) |
| MotH | 5′-AGGGACGGCTGACTTCTTCACC-3′ | S. typhimurium miaB (nt 1165–1144 in AJ249116) |
| PCR1 | 5′-AATACCGAGACGACGTTCGAGCTG-3′ | S. typhimurium f359a (nt 2376–2353 in AJ249116) |
| PCR2 | 5′-AAATCCGCACGTCAGGCTGGCTG-3′ | S. typhimurium o391 (nt 334–356 in AJ249116) |
| Sekv14 | 5′-TACTCGTTCATCTGACAGCC-3′ | S. typhimurium miaB (nt 683–664 in AJ249116) |
| T-I | 5′-GACAAGATGTGTATCCACCTTAAC-3′ | IS10R, facing outward |
| T-L | 5′-ACCTTTGGTCACCAACGCTTTTCC-3′ | Tn10dTc, ∼70 bp from the left end, facing outward |
| Tn10XbaI | 5′-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.