TABLE 1.
Strains, plasmids, and primers used in this study
| Strain, plasmid, or primer | Relevant characteristics | Reference or source |
|---|---|---|
| Serratia marcescens strains | ||
| CH-1 | Clinical isolate; does not contain spnTIR; swarming positive with flagellum | 18 |
| SS-1 | Environmental isolate; contains spnTIR; swarming negative without flagellum; sliding positive | 31 |
| SS-1ΔR | SS-1 spnR::Smr | 15 |
| Jun-1 | Isolated from repeated subcultures of SS-1; two TnTIR copies are in the chromosome | This study |
| Escherichia coli strains | ||
| K-12 MG1655 | Wild type | American Type Culture Collection (ATCC 47076) |
| CC118 | λ-pir lysogen [Δ(ara-leu) araD ΔlacX74 galE galK phoA20 thi-1 rpsE rpoB argE(Am) recA1]; permissive host for suicide plasmids requiring Pir protein | 14 |
| S17-1 | λ-pir lysogen [thi pro hsdR hsdM+recA RP4 2-Tc::Mu-Km::Tn7 (Tpr Smr)]; permissive host able to transfer suicide plasmid pDM4 to recipient cells via conjugation | 14 |
| Chromobacterium violaceum CV026 | Double mini-Tn5 mutant derived from C. violaceum; AHL biosensor Hgr; cvil::Tn5xyIE; Kanr; spontaneous Smr; non-AHL producer; nonpigmented unless exogenous AHL provided | 21 |
| Plasmids | ||
| pBAD18-Kan | pBAD18, arabinose regulation; Kanr | 13 |
| pDM4-230 | Derived from pDM4(23); contains sacBR genes, Cmr gene, and a 230-bp random sequence; requires Pir protein for replication | This study |
| pUT-Sm | Suicide plasmid containing mini-Tn5 (Smr); requires Pir protein for replication | 6 |
| pKOT101 | Derived from pUT; contains partial spnT coding region; Smr Kanr | This study |
| pKOT102 | Derived from pUT; contains araC and partial spnT coding region; PBAD promoter; Smr | This study |
| pKOT103 | Derived from pUT; contains araC and partial spnT coding region; PBAD promoter; Smr | This study |
| pJR203 | pBAD18-Kan::spnR | This study |
| Primers | ||
| TnP1 | 5′-GGGGCATCCGCTTCGCTTTCAGCAGCGTGAATGA-3′ | This study |
| TnP2 | 5′-GGGGTAATGGGAGCGAAGCTACAATATGACGACA-3′ | This study |
| XSF | 5′-GCCCACGCGCTCCCCGA-3′ | This study |
| TII | 5′-AGCGAGTGCATCATCGG-3′ | This study |
| XgeF | 5′-CCCGGTGTTTCATACAGCCAGCCC-3′ | This study |
| PsiI | 5′-GCATGCCATCGTGTATGGTCAGGGGC-3′ | This study |
| pBADF | 5′-CAAACCCTATGCTACTCCGTCAA-3′ | This study |
| pBADR | 5′-AATTCGCTAGCCCAAAAAAACG-3′ | This study |
| B6 | 5′-CTGCTGACCGGGCCTTCCACT-3′ | This study |
| FR-1 | 5′-CGGCCCTTGTCGGTCATAC-3′ | This study |