TABLE 1.
Plasmids and strains used in this studya
| Plasmid or strain | Relevant characteristic(s) | Source or reference |
|---|---|---|
| Plasmids | ||
| pCR-Blunt II-TOPO | E. coli cloning vector with ccd gene, Kmr + Zeor selection, blue-white screening; 3.5 kb | Invitrogen |
| pAMT1 | E coli-PAB shuttle vector Apr in E. coli and Cmr in PAB, derived from pUC18, pLME108 and cml/cmx PCR product; 6.3 kb | 4 |
| pTD104 | P4E::pctA promoter-gene fusion cloned in pAMT1; 6.9 kb | 4 |
| pTD10 | PAB cloning vector, Cmr in PAB, consisting of pLME108 and cml/cmxA PCR product derived from pAMT1; 3.6 kb | This study |
| pSL101 | PpampE::pcfI promoter-gene fusion cloned in pCR-Blunt II-TOPO; 4.4 kb | This study |
| pSL102 | pTD10 cloned as XbaI fragment into SpeI site of pSL101; 8.0 kb | This study |
| pSL103 | P. freudenreichii food-grade cloning vector, PpampE::pcfI propionicin F resistance marker, derived from pSL102; 3.1 kb | This study |
| pSL104 | P. freudenreichii food-grade cloning vector, propionicin F resistance from PpampS::pcfI, derived from pSL102; 2.7 kb | This study (see Fig. 3) |
| pSL106 | P4E::pctA promoter-gene fusion cloned in BamHI site of pSL104; 3.3 kb | This study |
| Strains | ||
| E. coli Top Ten | F−mcrA Δ(mrr-hsdRMS-mcrBC) Φ80lacZΔM15 ΔlacX74 recA1 araD139 Δ(araleu)7697 galU galK rpsL (Strr) endA1 nupG | Invitrogen |
| E. coli JM109 | F′ traD36 proA+ proB+ lacIq Δ(lacZ)M15/Δ(lac-proAB) glnV44 e14−gyrA96 recA1 relA1 endA1 thi hsdR17 | New England Biolabs |
| P. freudenreichii LMGT 2946 | Propionicin F producer | 3 |
| P. freudenreichii IFO12426 | High frequency of DNA transformation by electroporation | 4; IFO |
| P. thoenii 419b | Propionicin T1 producer | 10 |
| P. acidipropionici ATCC 4965 | Highly sensitive to propionicin T1 | ATCC |
| P. jensenii LMGT 3032 | pamA+ | 10 |
a
Abbreviations: ATCC, American Type Culture Collection (Rockville, MD); LMGT, Laboratory of Microbial Gene Technology, Ås, Norway; IFO, Institute of Fermentation Osaka, Japan; Cmr, chloramphenicol selection; Apr, ampicillin selection; Kmr, kanamycin selection; Zeor, zeomycin selection.
b
P. thoenii 419 was from the Environmental Bacteriology Culture Collection, University of the Orange Free State, Bloemfontein, Republic of South Africa.