Table 1.
Bacterial strains, plasmids, and primers used in this study.
| Bacterial strain, plasmid, or primer | Description or sequence | Source or reference |
|---|---|---|
| Bacterial strains | ||
| E. coli DH5α | Competent cells | Standard laboratory strain |
| L. monocytogenes | ||
| F2365 | Wildtype serotype 4b strain | Nelson et al. (2004) |
| F2365ΔccpC | F2365ΔccpC mutant strain | This study |
| F2365ΔccpC::pPL2-ccpC | F2365ΔccpC::pPL2-ccpC complement strain | This study |
| F2365Δhly | F2365Δhly mutant strain | Portnoy lab |
| F2365ΔplcA | F2365ΔplcA mutant strain | Portnoy lab |
| Plasmid | ||
| pHoss1 | 8,995 bp, pMAD,::secY antisense, ΔbgaB, Ampr, Eryr | Abdelhamed et al. (2015) |
| pPL2 | 6,123 bp, PSA attPP, Chlr | Lauer et al. (2002) |
| PccpCplasmid | pHoss,::ΔccpC, Ampr, Eryr | Abdelhamed et al. (2015) |
| pPL2-ccpC | pPL2,::ΔccpC, Ampr, Eryr | This study |
| Primers used for construction ΔccpC strain (5′-3′) | ||
| ccpC-A | aaaggggaattcCCACGCGCGTATAATAGCAA | EcoRI |
| ccpC-B | TTTCCTCCACCTGCAAATGA | |
| ccpC-C | TCATTTGCAGGTGGAGGAAAAACAGAACGTATCGGAAGGG | |
| ccpC-D | aaaggggtcgacCAAAATCCCCGTACGCCATT | SalI |
| ccpC-Seq | GCAAACGGATTTCCCAGTAA | |
| Primers used for construction complemented strain | ||
| ccpC-CompF1 | aaattcgagctcGCAGGTGGAGGAAAATAGATG | SacI |
| ccpC-CompR1 | aaattcgtcgacATTTTGCCATAGCTATTTAACTTGTTC | SalI |
| Primers for RT-qPCR (5′-3′) | ||
| plcA-F | GCATCACTTTCAGGCGTATTAG | |
| plcA-R | CGTGTCAGTTCTGGGAGTAG | |
| plcB- F | GCTTGACCGCAAGTGTTCTA | |
| plcB- R | GATTATCCGCGGACCAACTAAG | |
| gutM-F | GTTGCGGATAAATATGCTGAGAAA | |
| gutM-R | TGATGATAGTTGGTGAAAGTCTTGA | |
| inlB-F | GATGCGCTTCCTGCTTTAGA | |
| inlB-R | GAAAGTCCAGCATCCTCCATATT | |
| inlA_F | CGGCAAAGAAACAACCAAAGA | |
| inlA_R | GCATCAAACCACCCAACAAA | |
| recA-F | AGGCGAGCTTGTTGATATGG | |
| recA-R | CTTCCGTCGATTTCGTACTCTT | |
| lexA-F | GAAGCAGAGACACCCAATGT | |
| lexA-R | TCATACTTTCGCCGTCGATTT | |
| dnaA-F | CCTAGTTACGACACATGGATGAA | |
| dnaA-R | TCCTGCTCGCCATCAATAAA | |
| lspA-F | TGCCAAAGGAAAGCGACTAT | |
| lspA-R | GAGCATTAGTACGACACCAACT | |
| fur-F | CGGTGTTTCTCGGTATGACTT | |
| fur-R | GTCTGCAATCTGTGCAAATCC | |
| 16S rRNA-F | CAAGCGTTGTCCGGATTTATTG | |
| 16S rRNA-R | GCACTCCAGTCTTCCAGTTT | |
| LLO-F | CAAATGTGCCGCCAAGAAA | |
| LLO-R | CGAGAGCACCTGGATATGTTAG | |
| Crp/fnr-F | TAGGCGCAACCAACAGATT | |
| Crp/fnr-R | GTAAGCGGCCCGATACATT | |
| actA-F | GAAACAGCACCTTCGCTAGA | |
| actA-R | CTCTCCCGTTCAACTCTTCTTC | |
Underlined text denotes primer overlap with the ccpC-B primer, and bold faces in the primer sequences above indicate restriction enzyme sites. Ampr, ampicillin resistant; Eryr, erythromycin resistant; Chlr, chloramphenicol resistant.