Table 1.

Bacterial strains, plasmids, and oligonucleotides used in this study

Strain, plasmid, or oligonucleotideRelevant characteristics, construction, or nucleotide sequence (5′ → 3′)aReference or source
Strains
B. lactis DSM 10140Wild type22
E. coli XL1-Blue recA1 lac endA1 gyrA96 thi hsdR17 supE44 relA1 (F′ proAB lacI q lacZΔM15 Tn10Tetr)3
E. coliBL21(DE3)/pLysSF ompT hsdSB (rB mB ) gal dcm (DE3)/pLysS (Cmr)Novagen
Plasmids
 pUC18Ampr LacZ′; cloning vector; 2.7 kb35
 pCL1920SpcrStrr; cloning vector; 4.6 kb19
 pGEM-T EasyAmpr LacZ′; annealed T in both 3′ ends after linearization with EcoRV; 3.0 kbPromega
 pET-28a(+)Kanr LacI; expression vector; 5.4 kbNovagen
 pFPK1Ampr; 2.62-kbBamHI-BamHI fragment from strain DSM 10140 in pUC18; 5.3 kbThis study
 pFPK2Ampr; 1.6-kb PCR-derived fragment from strain DSM 10140 in pGEM-T Easy; 4.6 kbThis study
 pFPK3Spcr Strr; 0.85-kb BamHI-SmaI fragment from pFPK2 insertion in pCL1920; 5.4 kbThis study
 pFPK4SpcrStrr; 2.62-kb BamHI-insertion from pFPK1 in theBamHI site of pFPK3; 8.1 kbThis study
 pFPK5Kanr; 2.5-kb PCR-derived fragment from strain DSM 10140 in pET-28a(+); 7.9 kbThis study
Oligonucleotidesb
 pk55′-GGIACICCITGGCARAAR-3′ (974–991)This study
 pk65′-ATATATATARTAYTTRTCCATICCIATIAT-3′ (1019–1039 reverse)c This study
 pk75′-CATGGCAGAAGCTGGATCGTCCGGT-3′ (981–1005)This study
 pk95′-GCGAGATCCCGTGGCGT-3′ (2526–2542)This study
 pk155′-ATATATGAATTCATGACTAATCCTGTTATTGGTACC-3′ (956–979)c This study
 pk165′-AATTACAAGCTTTCACTCGTTGTCGCCGGCGG-3′ (3414–3433 reverse)c This study
  • a Position in the nucleotide sequence according to the numbering in the GenBank database.

  • b Oligonucleotides were synthesized by Microsynth AG, Balgach, Switzerland.

  • c The underlined sequences correspond to an introduced tail for cloning purposes.