TABLE 2.
Plasmids used in this study
| Plasmid | Description | Reference |
|---|---|---|
| pSC101-BAD-ETgA-tet | Plasmid used for Red/ET recombineering, tetracycline resistance | 55 |
| p15A-cm-tetR-tetO-hyg-ccdB | Plasmid used to amplify linear vector in Red/ET recombineering, chloramphenicol resistance | 55 |
| P15A-cm-tetR-tetO | Plasmid used as p15A empty vector without hyg and ccdB | This study |
| pBAD24Amp | Plasmid used to amplify bla gene in Red recombineering | Lab stock |
| pDS132kan | Suicide plasmid to construct in-frame deletions, kanamycin resistance | Lab stock |
| pDS132-ΔvasK | Suicide vector to construct in-frame deletion of vasK | 24 |
| pDS132-tseIHFH-AAA | Suicide vector to construct chromosomal tseIHFH-AAA | 36 |
| pDS132-tsePE663A | Suicide vector to construct chromosomal tsePE663A | 43 |
| pDS132-VipA-KI-sfGFP | Suicide vector to construct insertion of sfGFP | This study |
| pDS132-VipA-KI-N3-sfGFP | Suicide vector to construct insertion of N3 (noncontractile sheath mutation) and sfGFP | This study |
| pDS132-KI-NheI-F-NP | Suicide vector to construct insertion of NheI site in front of native promoter of hcp1 | This study |
| pDS132-KI-NheI-F-Tet | Suicide vector to construct insertion of NheI site in front of RBS of hcp1 | This study |
| pDS132-KI-NheI-B | Suicide vector to construct insertion of NheI site behind T6SS cluster | This study |
| pPSV37 | IPTG-inducible expression vector, gentamicin resistance | 81 |
| pPSV37-tsiI-3V5 | IPTG-inducible expression of TsiI with C-terminal 3V5 tag | This study |
| pBAD18 | Arabinose-inducible expression vector, kanamycin resistance | 82 |
| pBAD24-tseCtsiC | Arabinose-inducible expression of TseC and TsiC | This study |
| pBAD24-2404-2401 | Arabinose-inducible expression of genes from 2404 to 2401 | This study |
| pBAD24-2404-2401(TseCΔ14) | Arabinose-inducible expression of genes from 2404 to 2401 with TseCΔ14 mutant | This study |
| pT6SNP | Plasmid for expression of T6SS cluster under native promoter of hcp1, p15A-cm-tetR-tetO-hyg-ccdB backbone | This study |
| pT6SN3-NP | Plasmid for expression of T6SS cluster (noncontractile sheath mutation) under native promoter of hcp1, p15A-cm-tetR-tetO-hyg-ccdB backbone | This study |
| pΔvasKNP | Deletion of vasK (ΔvasK::ampr) in pT6SNP background | This study |
| pT6STet | Plasmid for expression of T6SS cluster under tetracycline-inducible promoter PtetO, p15A-cm-tetR-tetO-hyg-ccdB backbone | This study |
| pT6SN3-Tet | Plasmid for expression of T6SS cluster (noncontractile sheath mutation) under tetracycline-inducible promoter PtetO, p15A-cm-tetR-tetO-hyg-ccdB backbone | This study |
| pΔvasKTet | Deletion of vasK (ΔvasK::ampr) in pT6STet background | This study |
| pT6SNP_eff | Plasmid for expression of T6SS cluster (mutations of TseI catalytic residues H1507, F1508, and H1509 and TseP catalytic residue E663) under native promoter of hcp1, p15A-cm-tetR-tetO-hyg-ccdB backbone | This study |
| pT6SN3-NP_eff | Plasmid for expression of T6SS cluster (mutations of TseI catalytic residues H1507, F1508, H1509, and TseP catalytic residue E663; noncontractile sheath mutation) under native promoter of hcp1, p15A-cm-tetR-tetO-hyg-ccdB backbone | This study |
| pT6STet_eff | Plasmid for expression of T6SS cluster (mutations of TseI catalytic residues H1507, F1508, and H1509 and TseP catalytic residue E663) under tetracycline-inducible promoter PtetO, p15A-cm-tetR-tetO-hyg-ccdB backbone | This study |
| pT6SN3-Tet_eff | Plasmid for expression of T6SS cluster (mutations of TseI catalytic residues H1507, F1508, and H1509 and TseP catalytic residue E663; noncontractile sheath mutation) under tetracycline-inducible promoter PtetO, p15A-cm-tetR-tetO-hyg-ccdB backbone | This study |