TABLE 3.
Strains and plasmids used in this study
| Strain or plasmid | Relevant characteristics | Source or reference |
|---|---|---|
| Strains | ||
| E. coli JM109 | recA1 supE44 endA1 hsdR17 gyrA96 relA1 thi | 46 |
| E. coli JM110 | dam dcm mutant of JM109 | Novagen |
| H. mediterranei strains | ||
| DF50 | pyrF-deleted mutant of wild-type strain H. mediterranei ATCC 33500 | 47 |
| ΔEPS | EPS synthesis gene cluster deletion mutant of DF50 | 37 |
| Δpyk | pyk deletion mutant of DF50 | This study |
| Δpps strain | pps deletion mutant of DF50 | This study |
| Δpps-like gene strain | pps-like gene deletion mutant of DF50 | This study |
| Δpps Δpps-like gene strain | pps and pps-like gene deletion mutant of DF50 | This study |
| ΔEPS Δpps | pps deletion mutant of ΔEPS | This study |
| ΔEPS Δpps-like gene | pps-like gene deletion mutant of ΔEPS | This study |
| ΔEPS Δpps Δpps-like gene | pps and pps-like gene deletion mutant of ΔEPS | This study |
| H. volcanii H1424 | ΔpyrE2 ΔhdrB pitANph Δmrr cdc48d-Ct | 54 |
| Plasmids | ||
| pHFX | 4.0 kb; integration vector containing pyrF and its native promoter, Ampr | 47 |
| pHFX-pyk | 5.2 kb; integration vector of pHFX for pyk deletion | This study |
| pHFX-pps | 5.2 kb; integration vector of pHFX for pps deletion | This study |
| pHFX-pps-like gene | 5.2 kb; integration vector of pHFX for pps-like gene deletion | This study |
| pWL502 | 7.9 kb; shuttle vector with pyrF marker, Ampr | |
| pPYK | 9.6 kb; expression vector of pWL502 containing pyk | This study |
| pPPS | 10.1 kb; expression vector of pWL502 containing pps | This study |
| pPPS-like | 10.5 kb; expression vector of pWL502 containing pps-like gene | This study |
| pTA03 | 8.0 kb; expression vector with C-terminal His6 tag and promoter PphaR | 49 |
| pTA03-PYK | 9.7 kb; expression plasmid of pTA03 containing pyk | This study |
| pTA03-PPS | 10.2 kb; expression plasmid of pTA03 containing pps | This study |
| pTA03-mutant PYK | 9.7 kb; expression plasmid of pTA03 containing mutant pyk | This study |
| pTA03-mutant PPS | 10.2 kb; expression plasmid of pTA03 containing mutant pps | This study |
| pTA06 | 8.0 kb; expression vector with N-terminal His6 tag and promoter PphaR | 49 |
| pTA06-PPS-like | 10.6 kb; expression plasmid of pTA06 containing pps-like gene | This study |