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Table 1 Strains and plasmids used in this study a

From: Conserved active site cysteine residue of archaeal THI4 homolog is essential for thiamine biosynthesis in Haloferax volcanii

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 P fdx -pyrE2 with MCS [9]
pJAM202c Apr; Nvr; Hfx. volcanii-E. coli shuttle plasmid, empty vector control [12]
pJAM809 Apr; Nvr; pJAM202c carries P2 rrnA -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 P2 rrnA -hvo_0665-strepII (which replaced hvo_1862-strepII) This study
pJAM2822 Apr; Nvr; pJAM2821-derived carries P2 rrnA -hvo_0665 C165A-strepII This study
  1. 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.