TABLE 1.
Bacterial strains and plasmids used in this work
| Strain or plasmid | Relevant genotype or phenotype | Reference or source |
|---|---|---|
| R. leguminosarum | ||
| UPM791 | 128C53 wild type; Strr Nod+ Fix+ Hup+ | 32 |
| UPM1155 | UPM791 (Δhup/hyp cluster) Hup− | This lab |
| SM61 | UPM791 tatBC mutant | 25 |
| Escherichia coli | ||
| DH5α | recA1 endA1 gyrA96 thi hsdR17 supE44 relA1Δ(lacZYA-argF)U169 φ80dlacZΔM15 | 14 |
| S17.1 | thi pro hsdR−hsdM+recA RP4 2-Tc::Mu-Km::Tn7 (Spr Smr) | 36 |
| Plasmids | ||
| pALPF1 | pAL618 with P1 promoter replaced by fixN promoter | 6 |
| pALPF15 | pALPF1 ΔhupS | This work |
| pALPF2 | pALPF1 ΔhupL | This work |
| pALPF4 | pALPF1 ΔhupD | This work |
| pALPF6 | pALPF1 ΔhupG | This work |
| pALPF7 | pALPF1 ΔhupH | This work |
| pALPF8 | pALPF1 ΔhupI | This work |
| pALPF9 | pALPF1 ΔhupJ | This work |
| pKD3 | Template plasmid harboring FLP-mediated excision sequence flanking Cmr gene | 9 |
| pKD13 | Template plasmid harboring FLP-mediated excision sequence flanking Kmr gene | 9 |
| pPM70 | pKD3 derivative plasmid containing Strep-tag II sequence for N terminus end fusion | This work |
| pALPF34 | pALPF1 derivative plasmid carrying hupHstrep gene | This work |
| pBBR1MCS-2 | Broad-host-range plasmid; Kmr mob+ | 17 |
| pPM1350 | pBBR1MCS2 derivative plasmid containing a DNA fragment harboring PfixN promoter from R. leguminosarum | This work |
| pPM125 | pBBR1MCS2 derivative plasmid containing the hupG, hupHstrep, and hupI genes in an EcoRI DNA fragment | This work |
| pPM164 | pPM1350 derivative plasmid containing an NdeI-XbaI fragment harboring hupI under the control of PfixN | This work |
| pPM165 | pPM1350 derivative plasmid containing an NdeI-XhoI fragment harboring hupK under the control of PfixN | This work |
| pPM166 | pPM1350 derivative plasmid containing an NdeI-XhoI fragment harboring hupGstrep, hupH, and hupI under the control of PfixN | This work |
| PCR2.1-TOPO | PCR cloning vector | Invitrogen |