TABLE 2.
Primers used in this study
| Function and primer | Sequence (5′–3′) | Target or source |
|---|---|---|
| Construction of pGIBLD001 (P32-comX) | ||
| ComXSDLLCup | AAAAGAGCTCAATTATGAAAAAGAGG | comX |
| ComXSDLLCdown | AAAACTGC AGTTAATCATCATCTCG | comX |
| pMGP32UpMfeI | ATATCAATTGGTCCTCGGGATAT GATAAG | P32-comX cassette from pGIBLD011 |
| pMGTerDown | GACTTTGAACCTCAACTCC | P32-comX cassette from pGIBLD011 |
| Construction of pGIFPT001 (PxylT-comX) | ||
| pGhxylTcomXmaI | GTGGATCCCCCGGGCTGCAGGGTAGCGCAGAACGAGATTCACTTG | PxylT |
| pxylTcomXrec | GATAGTAACTCCTTAATTTTTATTTGC | PxylT |
| pGhxlTorfXMGrec | GCAAATAAAAATTAAGGAGTTACTATCATGGCAATCGTTTCAGCAGAAAAATTCG | pGIBLD001 |
| pGhxlTorfXMGXmaI | CCTGCAGCCCGGGGGATCCAC | pGIBLD001 |
| Construction of the PcomGA-luxAB reporter strain | ||
| LuxLLCf1 | ATAGTCTCGAGTTTAAGCAATTGAATCGCTAG | PcomGA promoter and pGIBLD012 |
| LuxLLCr1 | GCAAAAAGTTTCCAAATTTCATACTAGAATATACGCAATTTG | PcomGA promoter |
| LuxLLCf2 | CAAATTGCGTATATTCTAGTATGAAATTTGGAAACTTTTTGC | luxAB |
| LuxLLCr2 | GCGAAAGGATCCCTATTAGGTATATTCCATGTGG | luxAB |
| P3pseudoLLC | GCTCCCTCGAGGGCGGCTCTGTTGGATTAATATATGG | pGIBLD012 |
| Sequencing of rpsL | ||
| RpsLUnivUP | ATGCCTACAATTAACCAAT | rpsL |
| RpsLUnivDN | CACCGTATTTAGAACGG | rpsL |
| Amplification of rpsL | ||
| LLCdacARpsL | AGTAGTATCAGCACTGACAGC | rpsL |
| LLCfusARpsL | ACACCTTTGTTCTTGAAGG | rpsL |
| Construction of the comEC disruption mutant | ||
| ComECLLCUp | AAAGAGCTCAAAATAAAAATGAAATTATGG | comEC |
| ComECLLCDown | AAAGCTAGCGGGAAAAAATTGTGAATTAC | comEC |
| CatUpSpeI | AAAAACTAGTGCAGTTTAAATTCGGTCCTCGG | P32-cat cassette from pNZ5319 |
| CatDownSpeI | AAAAACTAGTGTACAGTCGGCATTATCTCAT | P32-cat cassette from pNZ5319 |
| Amplification of P32-cat cassette for construction of mecA, clpC, clpE, clpP, and comX deletion mutants | ||
| fgt02Fcat | TCCTCGGGATATGATAAGATTAATAG | P32-cat cassette from pUC18cm |
| fgt02RVcat | TCTCATATTATAAAAGCCAGTCATTAG | P32-cat cassette from pUC18cm |
| Construction and validation of the mecA deletion mutant | ||
| fgt01FmecArec | CTTTAATGATGGAATGATTG | mecA |
| fgt01RVmecArec | CTATTAATCTTATCATATCCCGAGGATCCATATAACTATATGAAACC | mecA |
| fgt03FmecArec | CTAATGACTGGCTTTTATAATATGAGACTTAGAAAAATCTAAATATGGTTG | mecA |
| fgt03RVmecArec | GAAGATTTTTAATTTCAAGTGTAG | mecA |
| MecAKOF | TCAGTACCGAAAAACGAATG | mecA |
| MecAKORV | ATTTACCAGTTCCGTTAGG | mecA |
| Construction and validation of the clpC deletion mutant | ||
| ClpCUPF | CTTTGGGTTCTAATTTATC | clpC |
| ClpCUPRVRec | CTATTAATCTTATCATATCCCGAGGACGTTGGTGTATATTTTAC | clpC |
| ClpCDownFRec | CTAATGACTGGCTTTTATAATATGAGATAGAAATAAAGGAAAGGAC | clpC |
| ClpCDownRV | TTGCTTTAAGGATAGTTTC | clpC |
| ClpCFdiag | AGAAGCCAATAATGACGATG | clpC |
| ClpCRVdiag | AGAATTCTGATGATGCACAGTC | clpC |
| Construction and validation of the clpE deletion mutant | ||
| ClpEUPF | CAGAGGACAGTAATATTTTT | clpE |
| CP_clpEUPRVRec | CTATTAATCTTATCATATCCCGAGGACTCCTCTTAATTCTCAGTAAT | clpE |
| ClpEDNFRec | CTAATGACTGGCTTTTATAATATGAGAGTCAGTAAAATAGTATTAGTGACA | clpE |
| ClpEDNR | GATGCTGGAACAATATTT | clpE |
| ClpEFdiag | CAAGGAACAGTGGAGCTTTTA | clpE |
| ClpERVdiag | GTAATTGATCCTGTTGGAGTTG | clpE |
| Construction and validation of the clpP deletion mutant | ||
| ClpPUPF | AAAGGTTGAATTTTCTG | clpP |
| ClpPUPRVRec | CTATTAATCTTATCATATCCCGAGGAAGATATGGACTTAATTTAGG | clpP |
| ClpPDNFRec | CTAATGACTGGCTTTTATAATATGAGAAATAAGCAATAAAGTCCTAG | clpP |
| ClpPDNR | TCCTTTACAGTTTTTAGATG | clpP |
| ClpPFdiag | AGAGGAGTTGTTCAAGAAGAAAG | clpP |
| ClpPRVdiag | AAATTACTTGGAGTTAAGGCC | clpP |
| Construction and validation of the comX deletion mutant | ||
| ComXUPF | GAAAAACGAAATTCAAAC | comX |
| ComXUPRVRec | CTATTAATCTTATCATATCCCGAGGAGGGAATTCTATTATAATGTTG | comX |
| ComXDNFRec | CTAATGACTGGCTTTTATAATATGAGAAGTTAAAACGGATACATAAG | comX |
| ComXDNR | TTAAGAGATTAGCTAAACATC | comX |
| ComXRVdiag | AACAGCTCAACGATTCTTTC | comX |
| ComXFdiag | ATTCCTTAGAAAGGAGGTGATC | comX |