Table 1.
Plasmids and bacterial strains used in this study
| Strain or plasmid | Relevant characteristicsa | Reference or source |
|---|---|---|
| Bacterial strains | ||
| V. vulnificus | ||
| MO6-24/O | Wild type; clinical isolate; virulent | Ref. 65 |
| KK131 | MO6-24/O with ΔplpA | This study |
| ZW141 | MO6-24/O with ΔhlyU | This study |
| DI0201 | MO6-24/O with Δcrp | Ref. 49 |
| KK151 | MO6-24/O with ΔhlyUΔcrp | This study |
| MS023 | MO6-24/O with ΔpilD | Ref. 20 |
| E. coli | ||
| S17-1λpir | λ-pir lysogen; thi pro hsdR hsdM+ recA RP4–2 Tc:: Mu-Km::Tn7;Tpr Smr; host for π-requiring plasmids; conjugal donor | Ref. 55 |
| BL21 (DE3) | F− ompT hsdSB (rB− mB−) gal dcm (DE3) | Laboratory collection |
| Plasmids | ||
| pDM4 | R6K γ ori sacB; suicide vector; oriT of RP4; Cmr | Ref. 54 |
| pKK1301 | pDM4 with ΔplpA; Cmr | This study |
| pZW1401 | pDM4 with ΔhlyU; Cmr | This study |
| pBS0907 | pDM4 with Δcrp; Cmr | Ref. 56 |
| pRK415 | Broad-host-range vector; IncP ori oriT RK2; Tcr | Ref. 57 |
| pKK1320 | pRK415 with plpA; Tcr | This study |
| pJH0311 | Broad-host-range vector; Apr, Cmr | Ref. 58 |
| pZW1510 | pJH0311 with hlyU; Apr, Cmr | This study |
| pKK1502 | pJH0311 with crp; Apr, Cmr | This study |
| pMS0908 | pJH0311 with pilD; Apr, Cmr | Ref. 20 |
| pMBP parallel 1 | MBP tag fusion expression vector; Apr | Ref. 59 |
| pKK1503 | pMBP parallel 1 with plpA-his6; Apr | This study |
| pET28a(+) | His6 tag fusion expression vector; Kmr | Novagen |
| pYU1317 | pET28a (+) with hlyU; Kmr | This study |
| pKK1504 | pET28a (+) with truncated plpA; Kmr | This study |
| pHK0201 | pRSET A with crp; Apr | Ref. 49 |
| pGEM-T Easy | PCR product cloning vector; Apr | Promega |
| pKK1505 | pGEM-T Easy with 461-bp fragment of plpA upstream region; Apr | This study |
| pBBR-lux | Broad-host-range vector with luxCDABE operon; Cmr | Ref. 64 |
| pKK1514 | pBBR-lux with 378-bp fragment of plpA upstream region; Cmr | This study |
| pKK1515 | pBBR-lux with 308-bp fragment of plpA upstream region; Cmr | This study |
| pKK1516 | pBBR-lux with 228-bp fragment of plpA upstream region; Cmr | This study |
| pKK1517 | pBBR-lux with 129-bp fragment of plpA upstream region; Cmr | This study |
a Tpr, trimethoprim-resistant; Smr, streptomycin-resistant; Cmr, chloramphenicol-resistant; Tcr, tetracycline-resistance; Apr, ampicillin-resistant; Kmr, kanamycin-resistant.