TABLE 1.
Bacterial strains and plasmidsa
| Strain or plasmid | Genotype or relevant characteristic(s) | Source or reference |
|---|---|---|
| E. coli | ||
| DH5α | recA1 endA1 gyrA96 thi hsdRl7 supE44 relA1 Δ(lacZYA-argF)U169 (φ80 lacZΔM15) deoR phoA | 27; Bethesda Research Laboratories |
| S17.1 | thi pro hsdR hsdM+ recA RP4::2-Tc::Mu-Kan::T7; Spr Smr | 46 |
| BW25113 | lacIqrrnBT14 ΔlacZWJ16 ΔhsdR514 ΔaraBADAH33 ΔrhaBADLD78 | 16 |
| MC4100 | F−araD139 Δ(argF-lac)U169 ptsF25 relA1 flb5301 rpsL150 | 12 |
| HYD723 | MC4100; nikA::MudI (Aprlac) | 51 |
| R. leguminosarum bv. viciae | ||
| UPM791 | 128C53 Strr; Nod+ Fix+ Hup+ | 30 |
| SPF25 | UPM791 with PfixN::hupSL | 10 |
| SPF2714 | SPF25 ΔhupE | This work |
| SPF228 | SPF25 ΔhupE2 | This work |
| SPF22A | SPF25 ΔhupE ΔhupE2 | This work |
| Rhizobium tropici CIAT899 | Wild type; Nod+ Fix+ Hup− | P. van Berkum (USDA, Beltsville, MD) |
| Rhizobium etli CE3 | CFN42; Nod+ Fix+ Hup−; Kanr | 36 |
| Mesorhizobium loti MAFF303099 | Wild type; Nod+ Fix+ Hup− | NIAS (Japan) |
| Plasmids | ||
| pCR2.1-TOPO | PCR product cloning vector; Apr Kanr | Invitrogenb |
| pRLH41 | pBluescriptSK containing a 5-kb DNA region of the UPM791 hup gene cluster | 31 |
| pAL618 | Cosmid pLAFR1 containing the UPM791 hup cluster; Tcr | 30 |
| pALPF1 | pAL618 with PfixN::hupSL | 10 |
| pALPFE | pALPF1 ΔhupE | This work |
| pK18mobsacB | pK18 derivative sacB; Kanr | 45 |
| pKPF1 | pK18mobsacB with PfixN::hupSL | 10 |
| pKE1 | pK18mobsacB with a deletion in hupE | This work |
| pKE2 | pK18mobsacB with a deletion in hupE2 | This work |
| pThupE2 | pCR2.1-TOPO with a 1.6-kb DNA of the hupE2 region | This work |
| pKD4 | Template plasmid harboring an FLP-mediated excision sequence flanking the Kanr gene | 16 |
| pCP20 | Plasmid for thermal induction of FLP synthesis; Cmr Apr | 16 |
| pBAD18-Kan | Expression vector carrying the arabinose PBAD promoter; Kanr | 26 |
| pBADE1 | pBAD18-Kan with the hupE gene | This work |
| pBADE2 | pBAD18-Kan with the hupE2 gene | This work |
| pBADE1.H series | pBADE1 with hupE mutated versions | This work |
a
Abbreviations: Ap, ampicillin; Kan, kanamycin; Str, streptomycin; Tc, tetracycline; Cm, chloramphenicol.