Table 1.
Strains and plasmids used in this study a
| Strain, plasmid or primer | Description | Source or reference |
|---|---|---|
| E. coli | ||
| TOP10 | F− recA1 endA1 hsdR17(rK − mK +) supE44 thi-1 gyrA relA1 | Invitrogen |
| GM2163 | F− ara-14 leuB6 fhuA31 lacY1 tsx78 glnV44 galK2 galT22 mcrA dcm-6 hisG4 rfbD1 rpsL136 dam13::Tn9 xylA5 mtl-1 thi-1 mcrB1 hsdR2 | New England Biolabs |
| XL1-Blue | endA1 gyrA96(nal R) thi-1 recA1 relA1 lac glnV44 F'[::Tn10 proAB + lacI q Δ(lacZ)M15] hsdR17(rK − mK +) | Stratagene |
| Hfx. volcanii | ||
| DS70 | wild-type isolate DS2 cured of plasmid pHV2 | [10] |
| H26 | DS70 ΔpyrE2 | [9] |
| NC1011 | H26 Δthi4 | This study |
| HM1052 | H26 ΔubaA | [11] |
| Plasmids | ||
| pTA131 | Apr; pBluescript II carries Pfdx-pyrE2 with MCS | [9] |
| pJAM202c | Apr; Nvr; Hfx. volcanii-E. coli shuttle plasmid, empty vector control | [12] |
| pJAM809 | Apr; Nvr; pJAM202c carries P2rrnA-hvo_1862-strepII | [13] |
| pJAM2819 | Apr; pTA131-based plasmid for pre-deletion of hvo_0665 | This study |
| pJAM2820 | Apr; pTA131-based plasmid for Δhvo_0665 | This study |
| pJAM2821 | Apr; Nvr; pJAM809-derived carries P2rrnA-hvo_0665-strepII (which replaced hvo_1862-strepII) | This study |
| pJAM2822 | Apr; Nvr; pJAM2821-derived carries P2rrnA-hvo_0665 C165A-strepII | This study |
aAbbreviations: Apr, ampicillin resistance; Nvr, novobiocin resistance; −strepII, C-terminal StrepII-tag fusion coding sequence (preceded by a KpnI site that facilitated generation of the C-terminal StrepII fusion with GlyThr linker). HvThi4 corresponds to HVO_0665, the gene locus tag number of GI: 292654829.