TABLE 1.
Bacterial strains, plasmids and primers used in this study
| Strain, plasmid, or primer | Genotype, description, or sequence | Reference or source |
|---|---|---|
| Plasmids | ||
| pACΩGm | pACYC184 derivative carrying Ω fragment, Gmr | 7 |
| pMOB3 | Kmr Cmr, 5% sucrose sensitive | 8 |
| pGEM-T Easy | Apr and TA cloning vector | Promega |
| pMT6121 | oriT, sacB, and Hgr Kmr in Tn1737KH | M. Tsuda (14) |
| pTK01 | tcp in pGEM-T Easy | This study |
| pTKG01 | tcp::Gmr in pGEM-T Easy | This study |
| pTKM01 | tcp::Gmr and mob in pGEM-T Easy | This study |
| pCR2.1 | Apr Kmr and TA cloning vector | Invitrogen |
| pTKM02 | tcp and mob in pCR2.1 | This study |
| pTKM03 | tcp, orf2, and mob in pCR2.1 | This study |
| pTKM04 | tcp and mob in pCR2.1 | This study |
| Strains | ||
| P. aeruginosa | ||
| SM7 | Smr derivative of PAO1 | This study |
| SM21 | Smr derivative of PAO1 | This study |
| KMX78 | Kmr Hgr, SM7 derivative harboring pMT6121::Tn1737KH | This study |
| KMX7803 | SM7 tcp::Tn1737KH(Kmr Hgr) | This study |
| TKP011 | SM21 tcp::Gmr | This study |
| TKP02 | KMX7803 derivative harboring pTKM02 | This study |
| TKP03 | KMX7803 derivative harboring pTKM03 | This study |
| TKP04 | TKP011 derivative harboring pTKM04 | This study |
| E. coli | ||
| CT726 | DH1(pMT6121::Tn1737KH) | M. Tsuda (14) |
| S17-1 | thi endA recA hsdR with RP4-2Tc:: Mu-Km::Tn7 integrated in chromosome | 10 |
| Primers | ||
| galk-s | 5′-CGCAGAACAGGCAGCAGAGCGT TT-3′ | This study |
| galk-a | 5′-GCAGCAGAGGCGGATAAAAGT GCG-3′ | This study |
| PAO561-s1 | 5′-GTGAAGGAAAGCGAGCACAACA AGGCGT-3′ | This study |
| PAO561-a | 5′-CGGGGTTAGAATGCCTCGCTC CCA-3′ | This study |
| PAO561-s2 | 5′-GTGATTGCCACTGTCCCACTCCG CTCAT-3′ | This study |