TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Descriptiona | Source |
|---|---|---|
| E. coli strains | ||
| DH5α | hsdR recA lacZYAF80 lacZΔM15 | BRL |
| BL21(DE3) | F− dcm ompT hsdSB(rB− mB−) gal λ(DE3) | Laboratory collection |
| S17-1 | F− recA hsdR RP4-2 (Tc::Mu) (Km::Tn7) lysogenized with λpir phage | Laboratory collection |
| DH5α/pUCP18/pDN19lacΩ-PpigA | E. coli DH5α containing plasmids pUCP18 and pDN19lacΩ-PpigA | This study |
| DH5α/pUCP18/pDN19lacΩ-PflhDC | E. coli DH5α containing plasmids pUCP18 and pDN19lacΩ-PflhDC | This study |
| DH5α/pXW1908/pDN19lacΩ-PpigA | E. coli DH5α containing plasmids pXW1908 and pDN19lacΩ-PpigA | This study |
| DH5α/pXW1908/pDN19lacΩ-PflhDC | E. coli DH5α containing plasmids pXW1908 and pDN19lacΩ-PflhDC | This study |
| DH5α/pXW2005/pDN19lacΩ-PpigA | E. coli DH5α containing plasmids pXW2005 and pDN19lacΩ-PpigA | This study |
| S. marcescens strains | ||
| JNB5-1 | S. marcescens wild-type strain | 20 |
| SK68 | rcsB::Gmr mutant of JNB5-1, prodigiosin-hyperproducing mutant | This study |
| SK68/pXW1908 | Mutant SK68 containing plasmid pXW1908 | This study |
| SK68/pUCP18 | Mutant SK68 containing empty vector pUCP18 | This study |
| ΔRcsB | rcsB deletion mutant of S. marcescens JNB5-1 | This study |
| ΔRcsB/pXW1908 | Mutant ΔRcsB containing plasmid pXW1908 | This study |
| ΔRcsB/pUCP18 | Mutant ΔRcsB containing empty vector pUCP18 | This study |
| ΔRcsA | rcsA deletion mutant of S. marcescens JNB5-1 | This study |
| ΔRcsC | rcsC deletion mutant of S. marcescens JNB5-1 | This study |
| ΔRcsD | rcsD deletion mutant of S. marcescens JNB5-1 | This study |
| ΔRcsF | rcsF deletion mutant of S. marcescens JNB5-1 | This study |
| ΔIgaA | igaA deletion mutant of S. marcescens JNB5-1 | This study |
| JNB5-1ΔflhDC | flhDC deletion mutant of S. marcescens JNB5-1 | This study |
| SK68ΔflhDC | rcsB flhDC double mutant of S. marcescens JNB5-1 | This study |
| JNB5-1/pDN19lacΩ-PpigA | S. marcescens JNB5-1 containing plasmid pDN19lacΩ-PpigA | This study |
| SK68/pDN19lacΩ-PpigA | S. marcescens SK68 containing plasmid pDN19lacΩ-PpigA | This study |
| JNB5-1ΔflhDC/pDN19lacΩ-PpigA | S. marcescens JNB5-1ΔflhDC containing plasmid pDN19lacΩ-PpigA | This study |
| Plasmids | ||
| pRK2013Tn5G | Tn5G-carrying plasmid, Kmr Gmr | 67 |
| pMD18T | Cloning vector, 2,692 bp, Ampr, lacZ | TaKaRa |
| pET28a | E. coli expression vector, Kmr | Laboratory collection |
| pXW1907 | rcsB gene cloned in pET28a for expression of RcsB protein in E. coli BL21(DE3) | This study |
| pUCP18 | Broad-host-range shuttle vector, Ampr | 68 |
| pXW1908 | rcsB gene driven by Plac promoter cloned into pUCP18, Ampr | This study |
| pXW2005 | flhDC genes driven by Plac promoter cloned into pUCP18, Ampr | This study |
| pDN19lacΩ | Promoterless lacZ fusion vector, Spr Smr Tcr | 69 |
| pDN19lacΩ-ppigA | pig operon promoter cloned in pDN19lacΩ, Spr Smr Tcr | This study |
| pDN19lacΩ-PflhDC | flhDC genes promoter cloned in pDN19lacΩ, Spr Smr Tcr | This study |
| pUTKm | Tn5-based delivery plasmid with Kmr Ampr | 56 |
a
Gm, gentamycin; Km, kanamycin; Amp, ampicillin; Sp, spectinomycin; Sm, streptomycin; Tc, tetracycline.