TABLE 5 .
Strains and plasmids
| Strain or plasmid | Description | Reference or source |
|---|---|---|
| Strains | ||
| CB15N | Wild-type C. crescentus | 38 |
| CB15N ΔssrA | In-frame deletion of ssrA | 27 |
| CB15N CC1214::Ω | CC1214 replaced by omega cassette | This study |
| KCK426 | E. coli BL21(DE3) carrying pET28CC1214 | This study |
| KCK428 | CB15N CC1214::Ω with Kanr marker 22.4 kb downstream | This study |
| KCK429 | CB15N CC1215 replaced by cat gene in antisense orientation | This study |
| KCK430 | CB15N CC1215 replaced by cat gene in sense orientation | This study |
| KCK431 | CB15N ΔssrA CC1215 replaced by cat gene in antisense orientation | This study |
| KCK432 | CB15N ΔssrA CC1215 replaced by cat gene in sense orientation | This study |
| Plasmids | ||
| pML81 | pJS14-derived plasmid with xylose promoter and NdeI site | M. T. Laub, unpublished |
| pML81-His6-CC1214 | Expresses CC1214 from xylose promoter | This study |
| pBBR1-tet | pBBR1 plasmid with tetA cloned into NotI site | Gift from Camille Lochte, Pasteur Institute of Lille |
| pBBR-PBN | pBBR1-tet-derived plasmid with PstI, BamHI, and NdeI cloning sites. | This study |
| pCC1214 | Expresses CC1214 from xylose promoter | This study |
| pACC1214G30A | Expresses CC1214 active-site mutant (GAQ) from xylose promoter | This study |
| pSsrA | Expresses ssrA from its native promoter | 23 |
| pBGS18T(1349674–1350629) | pBGS18T containing targeting fragment from position 1349674 to 1350629 | 34 |
| pET28CC1214 | Expresses CC1214 from T7 promoter | This study |
| pDHFR | Expresses DHFR from T7 promoter | New England Biolabs |
| pMT425 | Plasmid with vanillate-inducible promoter, cat gene | 39 |