TABLE 1.
Strains and plasmids used
| Strain or plasmid | Relevant genotype and/or descriptiona | Reference or source |
|---|---|---|
| Strains | ||
| E. coli | ||
| DH5 | Cloning strain | Invitrogen |
| S17-1 | M294::RP4-2 (Tc::Mu)(Km::Tn7); for plasmid mobilization | 46 |
| C. crescentus | ||
| CJW27 | CB15N | 15 |
| CJW686 | CB15N pleC::pBGENTpleCmyfp | 34 |
| CJW816 | CB15N ΔpleC/pMR20 | P. Angelastro (gift) |
| CJW1208 | CB15N ΔcreS | 6 |
| CJW1298 | CB15N/pMR20 | P. Angelastro (gift) |
| CJW1406 | CB15N tipN::pBGENT{tipNgfp | 27 |
| CJW1524 | CB15N ΔfliM | This study |
| CJW1757 | CB15N ΔpflI | This study |
| CJW2179 | CB15N/pMR20pxylpflIgfp | This study |
| CJW2181 | CB15N pflI::pHL32{pflIgfp | This study |
| CJW2182 | CB15N ΔfliF pflI::pHL32{pflIgfp | This study |
| CJW2184 | CB15N ΔpflI pleC::pBGENTpleCmyfp | This study |
| CJW2185 | CB15N pleC::pBGENTpleCmyfp/pMR20pxylpflI | This study |
| CJW2186 | CB15N ΔpflI tipN::pBGENT{tipNgfp | This study |
| CJW2187 | CB15N tipN::pBGENT{tipNgfp/pMR20pxylpflI | This study |
| CJW2188 | CB15N ΔpflI cpaE::cpaE-yfp | This study |
| CJW2189 | CB15N cpaE::cpaE-yfp/pMR20pxylpflI | This study |
| CJW2192 | CB15N/pMR20pxylpflI | This study |
| LS 1218 | CB15N ΔfliF | 22 |
| PV418 | CB15N cpaE::cpaE-yfp | 49 |
| UJ506 | CB15N ΔpleC | 2 |
| Plasmids | ||
| pBGENT | Gmr variant of pBGST18 integration vector | 34 |
| pBGENT{tipNgfp | Integration plasmid with 3′ end of tipN fused to gfp | 27 |
| pBGENTpleC-myfp | Integration plasmid with full-length pleC fused to monomeric yfp | 34 |
| pBGENTpxylpflIgfp | Integration plasmid with pflI-gfp under the xylose-inducible promoter | This study |
| pBluescript KS(+) | Apr cloning vector | Stratagene |
| pEGFP-N2 | Kmr cloning vector harboring gfp | Clontech |
| pHL32 | Kmr variant of pBGST18 with pBluescript KS multiple cloning site | 27 |
| pHL32{pflIgfp | Integration plasmid with 3′ end of pflI fused to gfp | This study |
| pHPV227 (pNPTScpaEyfp) | Sucrose suicide vector harboring cpaE fused to yfp | 49 |
| pJS14 | Cmr medium-copy-no. pBBR1-derived broad-host-range vector | Jeff Skerker |
| pJS14pflIgfp | pJS14 carrying pflI fused to gfp | This study |
| pJS14pxylpflI | pJS14 carrying pflI under the control of the xylose-inducible promoter | This study |
| pJS14pxylpflIgfp | pJS14 carrying pflI fused to gfp under the control of the xylose-inducible promoter | This study |
| pMR20 | Tcr low-copy-no. broad-host-range vector | 44 |
| pMR20pxylpflI | pMR20 carrying pflI under the control of the xylose-inducible promoter | This study |
| pMR20pxylpflIgfp | pMR20 carrying pflI-gfp under the control of the xylose-inducible promoter | This study |
| pNPTS138 | Litmus 38-derived vector; oriT sacB Kmr | MRK Alley |
| pNPTSfliMko | Sucrose suicide vector with sequences upstream and downstream of fliM | This study |
| pNPTSpflIko | Sucrose suicide vector with sequences upstream and downstream of pflI | This study |
a
Tc, tetracycline; Km, kanamycin; Gm, gentamicin; Ap, ampicillin; Cm, chloramphenicol.