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Table 3 Bacterial strains and plasmids used in this work

From: Dual role of HupF in the biosynthesis of [NiFe] hydrogenase in Rhizobium leguminosarum

Strain or plasmid Relevant genotype or phenotype Source or reference
Rhizobium leguminosarum   
UPM791 128C53 wild type; Strr Nod+ Fix+ Hup+ [40]
UPM1155 UPM791 ( Δhup/hyp cluster) Hup- [19]
Escherichia coli   
DH5α recA1 endA1 gyrA96 thi hsdR17 supE44 relA1 Δ(lacZYA-argF)U169 Φ80dlacZΔM15 [41]
S17.1 thi pro hsdR-hsdM+recA RP4::2-Tc::Mu-Kan::T7; (Spr Smr) [42]
Plasmids   
pAL618 pLAFR1-based cosmid containing the whole R. leguminosarum hydrogenase gene cluster [40]
pALPF1 pAL618 with hupSL promoter replaced by fixN promoter (P fixN ) [18]
pALPF2 pALPF1 ΔhupL [19]
pALPF4 pALPF1 ΔhupD [19]
pALPF5 pALPF1 ΔhupF This work
pALPF10 pALPF1 ΔhupK This work
pALPF14 pALPF1 ΔhypC This work
pALPF382 pALPF1 derivative carrying hupF ST gene This work
pBBR1MCS-2 Broad-host-range plasmid; Kmr mob+ [43]
pKD3 Template plasmid harbouring FLP-mediated excision sequences flanking Cmr gene [44]
pPM71 PKD3 derivative containing Strep-tag II sequence for C-terminal end fusion This work
pPM1350 pBBR1MCS-2 derivative containing a DNA fragment harbouring P fixN promoter from R. leguminosarum [19]
pPM501 pPM1350 derivative containing an NdeI-XbaI fragment harbouring HupF ST under the control of PfixN This work
pPM501C pPM501 derivative containing a deletion of the 25 3′codons of hupF This work
pPCR2.1-TOPO PCR cloning vector Invitrogen