TABLE 2.
Plasmids and strains
| Plasmid or strain | Relevant characteristicsa | Reference or source |
|---|---|---|
| Plasmids | ||
| pET-22b(+) | Ampr; expression vector; T7 promoter, C-terminal 6×His tag | Novagen |
| pTA131 | Ampr; pyrE2+ marker | 32 |
| pTA963 | Ampr; pyrE2+ and hdrB+ markers; p.tnaA (Trp-inducible promoter) | 28 |
| pFH24 | pTA131 with insert: XhoI::700 bp 5′ of hvo_0405::700 bp 3′ of hvo_0405::XbaI | This study |
| pFH25 | pTA963 with insert: p.tnaA::NdeI::hvo_0405 cds::GGP linker::6×His::stop::EcoRI | This study |
| pJS143 | pET-22b(+) with insert: T7 promoter::lacO::ribosomal binding site::NdeI::hvo_0405 (bp 64–1344)::KL(HindIII)AAALE linker::6×His tag::stop | This study |
| pJS151 | pTA963 with insert: p.tnaA::Hvo_0405 cds with coding bases 7–12 mutated from CGCCGC to AAGAAG (changes RR to LL)::GGP linker::6×His tag::stop | This study |
| Strains | ||
| DH5α | E. coli F− ϕ80dlacZΔM15 Δ(lacZYA-argF)U169 recA1 endA1 hsdR17(rK− mK+) phoA supE44 thi-1 gyrA96 relA1 λ− | Invitrogen |
| DL739 | E. coli Cmr; MC4100 recA dam-13::Tn9 | 31 |
| BL21(DE3) | E. coli Cmr; strain B F− ompT gal dcm lon hsdSB(rB− mB−) gal dcm (DE3 λ[lacI lacUV5-T7p07 ind1 sam7 nin5]) [malB+]K-12(λS)(pLysS[T7p20 orip15A]) | 33 |
| H53 | H. volcanii DS70 ΔpyrE2 ΔtrpA | 32 |
| AF103 | H53 ΔartA | 12 |
| FH24 | H53 Δhvo_0405 | This study |
| FH35 | H53 ΔartA Δhvo_0405 | This study |
| JS1 | H53 containing pTA963 | This study |
| JS13 | H53 containing pFH25 | This study |
| JS16 | FH24 containing pTA963 | This study |
| JS17 | FH24 containing pFH25 | This study |
| JS23 | FH35 containing pTA963 | This study |
| FH41 | FH35 containing pFH25 | This study |
| FH59 | FH24 containing pJS151 | This study |
a
Ampr, ampicillin resistance; cds, coding sequence; Cmr, chloramphenicol resistance.