Table 1.

Bacterial strains and plasmids used in this study

Strain or plasmidOriginCoordinates of TP901-1 fragment(s)aAntibiotic resistanceReference or source
Strains
L. lactissubsp.   cremoris
  JM342MG1363 Δupp tdk23
  AB112JM342::pAB112−104–228ErmrThis study
  AB174aJM342::pAB174a−98–1910, 2521–2708ErmrThis study
E. coli DH5αø80lacZΔM15 Δ(lacZYA-argF)U169 recA1 endA1 hsdR17 supE44 thi-1 gyrA96 relA1Lab strain
Plasmids
 pCI3340Shuttle vector for E. coli and L. lactisCamr10
 pCI372Shuttle vector for E. coli and L. lactisCamr10
 pTRKL2Shuttle vector for E. coli and L. lactisErmr25
 pAK80Shuttle vector for E. coli and L. lactisErmr12
 pGEM-3Zf(+)E. colivectorAmprPromega
 pGEM-7Zf(+)E. coli vectorAmprPromega
 pIC-19RE. colivectorAmpr21
 pACYC184E. colivectorCamrNew England Biolabs
 pBF12pMOSblue-T::333-bp TP901-1attP−104–228Ampr, Ermr3
 pPM92pIC-19R::TP901-1orf492621–5476Ampr20
 pPM115pCI3340::TP901-1orf291287–5476Camr17
 pPM129pAK80::TP901-1orf472621–4672Ermr18
 pJM300pUN121::uppAmpr, Tetr22
 pAJ37pCI372::TP901-1 attP,orf12,orf46d−98–4475dCamrThis study
 pAJ39pCI372::TP901-1 attP,orf1,orf36e−98–4475eCamrThis study
 pAJ41pCI372::TP901-1 attP,orf26f−98–4475fCamrThis study
 pAJ43pCI372::TP901-1 attP,orf16−98–4475CamrThis study
 pAJ95pAK80::TP901-1pR-pL3142–3332Ermr14
 pAB35pCI372::TP901-1 attP,orf1,orf46−98–1910, 2521–4475CamrThis study
 pAB112pMOSblue-T::333-bp TP901-1attP, upp−104–228Ampr, ErmrThis study
 pAB174apGEM-7Zf(+)::TP901-1 attP,orf1,bupp−98–1910, 2521–2708Ampr, ErmrThis study
 pAB174bpGEM-7Zf(+)::TP901-1 attP,orf1,cupp−98–1910, 2521–2708Ampr, ErmrThis study
 pAB204pGEM-3ZF(+)::uppAmprThis study
 pAB211pIC-19R::uppAmprThis study
 pAB221pCI3340::TP901-1orf251287–3900CamrThis study
 pAB223pCI3340::TP901-1orf452621–3900CamrThis study
 pAB227pTRKL2::uppErmrThis study
 pAB235pCI372::TP901-1 attP,orf1−98–1910, 2521–2708CamrThis study
 pAB241pCI3340::TP901-1orf472621–4672CamrThis study
 pAB243pCI3340::TP901-1orf45,orf72621–3900, 4353–4672CamrThis study
 pAB244pCI3340::TP901-1orf74353–4672CamrThis study
 pAB245pCI3340::TP901-1pR-pL,orf73142–3332, 4353–4672CamrThis study
 pLB28pGEM-7Zf(+)::TP901-1 attP,orf23−98–2708fAmpr6
 pLB29pGEM-7Zf(+)::TP901-1 attP,orf13−98–2708Ampr6
 pLB44pGEM-7Zf(+)::TP901-1 attP,orf1c−98–1910, 2521–2708Ampr6
 pLB55pCI372::TP901-1 attP,orf12d−98–2708dCamrThis study
 pLB56pCI372::TP901-1 attP,orf1,orf3e−98–2708eCamrThis study
 pLB57pCI372::TP901-1 attP,orf23f−98–2708fCamrThis study
 pLB58pCI372::TP901-1 attP,orf13−98–2708CamrThis study
 pLB61pGEM-7Zf(+)::TP901-1orf125–1910Ampr3
 pLB71pGEM-7Zf(+)::TP901-1 attP,orf12d−98–2708dAmpr6
 pLB72pGEM-7Zf(+)::TP901-1 attP,orf1,orf3e−98–2708eAmpr6
 pLB81pACYC184::TP901-1orf125–1910CamrThis study
  • a Coordinates of the TP901-1 fragment(s), according to the numbering system introduced previously (19), where first base pair in the core region is assigned position 1 and the last base pair of the core is assigned position 5. Since the sequence of the entire phage genome is not known, we have assigned the bases on the other side of the core from orf1negative numbers. Positive coordinates thus correspond to the sequence reported under EMBL accession no. Y14232 . Negative coordinates −1 to −104 correspond to bases 2727 to 2831 in the sequence reported under EMBL accession no. X85213 .

  • b The erm gene is oriented divergently from orf1.

  • c The erm gene is oriented in the same direction as orf1.

  • d Contains an amber stop mutation inorf3.

  • e Contains an amber stop mutation inorf2.

  • f Contains an amber stop mutation inorf1.