Table 1.
Strains, plasmids, and primers used in the study. Restriction recognition sites are underlined.
| Strain, Plasmid, or Gene | Relevant Features or Sequence (5’–3’) | Ref. or Source |
|---|---|---|
| Strain | ||
| E. coli | ||
| DH5α | endA1 glnV44 thi-1 recA1 relA1 gyrA96 deoR F- Φ80dlacZΔM15 Δ(lacZYA-argF)U169, hsdR17(rK- mK+), λ– | Invitrogen |
| TOP10 | F– mcrA Δ(mrr-hsdRMS-mcrBC) Φ80lacZΔM15 ΔlacX74 recA1 araD139 Δ(ara leu) 7697 galU galK rpsL (StrR) endA1 nupG | Life technologies |
| BL21 λ(D3) | E. coli B F – dcm ompT hsdS (rB– mB–) gal λ(DE3) | [15] |
| L. lactis | ||
| NZ9000 | MG1363 nisRK ΔpepN | [23] |
| Plasmid | ||
| pNZ8148 | pSH71 derivative, PnisA, CmR, nisin-controlled expression | [23] |
| pSDBA3b | pNZ8148 containing gene fusion of Usp45 signal peptide, B domain, and cA | [24] |
| pET-REX009 | pET28b containing a fusion gene of REX009, tolA protein, and AviTag consensus | [9] |
| pET-REX115 | pET28b containing a fusion gene of REX115, tolA protein, and AviTag consensus | [9] |
| pET-REX125 | pET28b containing a fusion gene of REX125, tolA protein, and AviTag consensus | [9] |
| pSD-REX009 | pNZ8148 containing gene fusion of Usp45 signal peptide, REX009, and cA | This work |
| pSD-REX115 | pNZ8148 containing gene fusion of Usp45 signal peptide, REX115, and cA | This work |
| pSD-REX125 | pNZ8148 containing gene fusion of Usp45 signal peptide, REX125, and cA | This work |
| pSD-REX009-FLAG | pNZ8148 containing gene fusion of Usp45 signal peptide, FLAG tag, REX009, and cA | This work |
| pSD-REX115-FLAG | pNZ8148 containing gene fusion of Usp45 signal peptide, FLAG tag, REX115, and cA | This work |
| pSD-REX125-FLAG | pNZ8148 containing gene fusion of Usp45 signal peptide, FLAG tag, REX125, and cA | This work |
| pSC-REX009 | pNZ8148 containing gene fusion of Usp45 signal peptide and REX009 | This work |
| pSC-REX115 | pNZ8148 containing gene fusion of Usp45 signal peptide and REX115 | This work |
| pSC-REX125 | pNZ8148 containing gene fusion of Usp45 signal peptide and REX125 | This work |
| pSC-REX009-FLAG | pNZ8148 containing gene fusion of Usp45 signal peptide, FLAG tag, and REX009 | This work |
| pSC-REX115-FLAG | pNZ8148 containing gene fusion of Usp45 signal peptide, FLAG tag, and REX115 | This work |
| pSC-REX125-FLAG | pNZ8148 containing gene fusion of Usp45 signal peptide, FLAG tag, and REX125 | This work |
| Primer | ||
| ILP030-F | TGGATCCTTAGCTGAAGCTAAAGTC | This work |
| Rex009-R-Eco | AGAATTCAGGTAACGAAGCTAAAATC | This work |
| Rex009-R-Xba | ATCTAGAAGGTAACGAAGCTAAAATC | This work |
| Rex115-R-Eco | AGAATTCAAGGTAAAACAGCTAAAATCC | This work |
| Rex115-R-Xba | ATCTAGAAGGTAAAACAGCTAAAATCC | This work |
| Rex125-R-Eco | AGAATTCAAGGTAACGCAGCTAAAATAG | This work |
| Rex125-R-Xba | AGAATTCAGGTAACGCAGCTAAAATAG | This work |
| Usp1-NcoI | ATAACCATGGCTAAAAAAAAGATTATCTCAGCTATTTTAATG | [19] |
| FLAG_Bam_R | GGATCCTTTATCATCGTCGTCTTTATAATCAGCGTAAACACCTGACAACG | [25] |