TABLE 1.
Oligonucleotides used in this study
| Oligonucleotide | Sequence (5′→3′)a | Application |
|---|---|---|
| m-prfA-Nde | ATTGGGGGATGACATATGAACGC | Purification of untagged PrfA and PrfA* proteins of L. monocytogenes |
| m-prfA-Bgl | ATGAAAATGCTTCTGTAAGATCTTTATTCG | |
| i-prfA-Bam | TAAAAACGATTGGGGGATCCAAAATGGACG | Purification of His6-tagged PrfA of L. ivanovii (PrfALi) |
| i-prfA-Kpn | TCCTGTTTTTTTTGGTACCGTAGTATTTAATTG | |
| s-prfA-Bam | GGAGGATGACGATGGATCCACTAAAGC | Purification of His6-tagged PrfA of L. seeligeri (PrfALs) |
| s-prfA-Pst | ACAATTAATAAAAAGCGCTGCAGGGTC | |
| s-prfA-rkmb2 | TGAATTTTTATACACAATGACGTTTTCTTGTTTTAG | Purification of His6-tagged hybrid sm-PrfA |
| m-prfA-rkmb | CAAGAAAACGTCATTGTGTATAAAAATTCATGCTT | |
| m-prfA-Pst | TGAAAATGCTTCTGTAAGTTCTGCAGTCG | |
| m-actA1 | GAAGATGCTTCTAGAAAAGTTGCTGA | In vitro transcription assay with L. monocytogenes actA promoter (PactALm) |
| m-actA2 | CTCCCTCCTCGGAATTCGCTAA | |
| i-actA1 | ATGGGAAGAATTCGGCGTTAGATAAGGG | In vitro transcription assay with L. ivanovii actA promoter (PactALi) |
| i-actA2 | AATCTATCTAGGATCCCTTTTTTACTCC | |
| s-actA1 | GAAGTTGTCTAGAAAATTGCCGAAGCATGGC | In vitro transcription assay with L. seeligeri actA promoter (PactALs) |
| s-actA2 | GTTAATAGAAATCAGGATCCTAAAATCTACTGAC | |
| m-hly1 | CTGTTTTTAATTGACTGCAGTTTTCCGGC | In vitro transcription assay with L. monocytogenes hly promoter (PhlyLm) |
| m-hly2 | CAGTTATTAAGCGAATAAGCTTTTCCGCC | |
| i-hly1 | TGTAATAAAAACGAGCTCTATTTTTTTCATGG | In vitro transcription assay with L. ivanovii hly promoter (PhlyLi) |
| i-hly2 | TGTCATTAAAAGTAGAATTCTTTTTTTCATGG | |
| s-hly1 | TAGATGAAAAAGCTTTCCCGTGCATAGG | In vitro transcription assay with L. seeligeri hly promoter (PhlyLs) |
| s-hly2 | AACGACATGATATCTAGACCAAATATTTTC | |
| m-actA3 | GATGCTTCTAGAAAAGTTGCTGAAGC | EMSA with PactALm |
| m-actA4 | TATTCTAGAATTATTTTAAGAATATCA | |
| i-actA3 | CATGGGAAGAAGTTGGCGTTAGAT | EMSA with PactALi |
| i-actA4 | CAATTCATGAATTGCTTCAAGAATAGCA | |
| s-actA3 | CATGGCTCGGTGTAGGTATTTC | EMSA with PactALs |
| s-actA4 | ATTCATGAATTATTTACAGAATAACAC | |
| m-hly3 | TCCTATCTAGAAGTGACTTTTATGTT | EMSA with PhlyLm |
| m-hly4 | GCTTCTCTAGATGAAACGCAATATTA | |
| i-hly3 | GCCTATCTAGAAAATAATTTCACATC | EMSA with PhlyLi |
| i-hly4 | CTTCCAAGATTTAGAAACGCAATATTA | |
| s-hly3 | GTCTTATCTAGATCAACATTAACATA | EMSA with PhlyLs |
| s-hly4 | CGATTCATTAGCTTGAAACGCAATATTA | |
| SPR-hly1 | TGAGGCATTAACATTTGTTAACGACGAT | SPR with PrfA box of PhlyLm |
| SPR-hly2 | ATCGTCGTTAACAAATGTTAATGCCTCA | |
| SPR-ctrl1 | TGAGGCAATGGCATAGGCAACCGACGAT | SPR with nonspecific DNA for control |
| SPR-ctrl2 | ATCGTCGGTTGCCTATGCCATTGCCTCA | |
| prfA_Salup> | CAATAAAGTCGACTAACATATATTATTCCTAC | Allelic replacement of prfALm with prfALs in L. monocytogenes EGD-e (upstream region) |
| <prfA_Alwup | TCTTCTGCTTGAGTGCTCACGTCTCATCC | |
| s-prfA_bnt> | GTGAGCACTAAAGCAGAGGATTTTAAAGAATA | Allelic replacement of prfALm with prfALs in L. monocytogenes (coding region) |
| <s-prfA_Bse | TTATCCCAGCTATTATGACAAGCTAAG | |
| prfA_Bsedn> | GTCCTGCTAGCTGGGATAAATTTAATTAAATC | Allelic replacement of prfALm with prfALs in L. monocytogenes (downstream region) |
| <prfA_Bamdn | GCTAAAGGATCCCGTGAAGCTGGCGCAACAAA | |
| prfA_Salup> | CAATAAAGTCGACTAACATATATTATTCCTAC | Cloning of prfALm into L. monocytogenes EGD-e ΔprfA |
| <prfA_Bgldn | GCTAAAAGATCTCGTGAAGCTGGCGCAACAAA |
a
Introduced restriction sites are underlined; bold letters show overlapping sequences for recombinant PCR.