TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Relevant property or propertiesa | Source or reference |
|---|---|---|
| Strains | ||
| E. coli JM109 | Host for cloning vectors | Laboratory stock |
| E. coli JM110 | Host used for preparing plasmid that is free of Dam or Dcm methylation | Laboratory stock |
| Ha. japonica | Wild type (JCM 7785T) | 21 |
| Ha. japonica Δc0507 | c0507 gene mutant of Ha. japonica | This work |
| Ha. japonica Δc0506 | c0506 gene mutant of Ha. japonica | This work |
| Ha. japonica Δc0505 | c0505 gene mutant of Ha. japonica | This work |
| Ha. japonica Δc0507(pJc0507) | Δc0507 complement with pJc0507 | This work |
| Ha. japonica Δc0506(pJc0506) | Δc0506 complement with pJc0506 | This work |
| Ha. japonica Δc0505(pJc0505) | Δc0505 complement with pJc0505 | This work |
| Plasmids | ||
| pUC119 | E. coli cloning vector; Apr | Laboratory stock |
| pWL102 | E. coli-haloarchaea shuttle vector; Apr Mevr | 29 |
| pDrHj2b | E. coli-haloarchaea shuttle vector; Apr Mevr | Laboratory stock |
| pWL102-Δc0507 | pWL102 derivative containing disrupted fragment of c0507 gene | This work |
| pDrHj2-Δc0506 | pDrHj2 derivative containing disrupted fragment of c0506 gene | This work |
| pDrHj2-Δc0505 | pDrHj2 derivative containing disrupted fragment of c0505 gene | This work |
| pJFZ33 | pWL102 derivative carrying Ha. japonica csg promoter and ftsZ2 gene; Apr Mevr | 27 |
| pJc0507 | pJFZ33 derivative in which ftsZ2 is replaced by the c0507 gene | This work |
| pJc0506 | pJFZ33 derivative in which ftsZ2 is replaced by the c0506 gene | This work |
| pJc0505 | pJFZ33 derivative in which ftsZ2 is replaced by the c0505 gene | This work |
a
Apr, ampicillin resistance; Mevr, pravastatin resistance.
b
The pDrHj2 plasmid was constructed as follows. The sequence between SacI and KpnI in pUC119 was replaced with the mevinoline resistance gene, and the sequence between HindIII and XbaI was replaced with the gene of a halophilic β-galactosidase (bgaH) under the control of csg promoter. The bgaH gene was obtained from Haloferax alicantei and was oriented in the direction opposite that in which the mevinoline resistance gene was oriented. The csg promoter is from Haloarcula japonica. Ha. japonica has a large amount of a glycoprotein (cell surface glycoprotein [CSG]) on the cell surface, suggesting that the csg promoter is powerful.