Table 1.
Bacterial strains, plasmids and oligonucleotides used in this study.
| Strain, Plasmid or Oligonucleotide | Characteristics or Sequence a | Reference or Source |
|---|---|---|
| Bacterial strains | ||
| Bradyrhizobium elkanii | ||
| USDA61 | Wild-type strain, Polr | Keyser b |
| BErhcJ | USDA61 derivative harboring insertion in rhcJ region, Polr, Kmr, Tcr | [12] |
| BE5 | Tn5 mutant of USDA61, Polr, Kmr | This study |
| BE53 | Tn5 mutant of USDA61, Polr, Kmr | This study |
| BE85 | Tn5 mutant of USDA61, Polr, Kmr | This study |
| BE103 | Tn5 mutant of USDA61, Polr, Kmr | This study |
| BE168 | Tn5 mutant of USDA61, Polr, Kmr | This study |
| BE53S | USDA61 derivative containing an insertion of plasmid pSUPSCAKm:int innB, Polr, Kmr | This study |
| Escherichia coli | ||
| HB101 | recA, hsdR, hsdM, pro, Smr | Invitrogen, Carlsbad, CA, USA |
| S17-1 | pro recA RP4-2(Tcs:Mu) (Kms:Tn7); Mob+ | [20] |
| DH10B | Cloning strain | Invitrogen |
| Plasmids | ||
| pUTKm | Transposon delivery vector; Apr, Kmr | [21] |
| pRK2013 | Helper plasmid, ColE1 replicon carrying RK2 transfer genes; Kmr, tra | [22] |
| pSUPSCAKm | Derivative of pSUPPOL2SCA [23] with a kanamycin resistance gene in the DraI site, oriT of RP4, Tcr, Kmr | This study |
| pSUPSCAKm:int innB | pSUPSCAKm carrying a 0.5-kb DNA fragment containing internal sequence of innB, Kmr | This study |
| Oligonucleotides | ||
| Linker 1 | 5′-TTTCTGCTCGAATTCAAGCTTCTAACGATGTACGGGGACACATG-3′ | [24] |
| Linker 2 | 5′-TGTCCCCGTACATCGTTAGAACTACTCGTACCATCCACAT-3′ | [24] |
| Y-linker primer | 5′-CTGCTCGAATTCAAGCTTCT-3′ | [24] |
| Tn5 primer | 5′-GGCCAGATCTGATCAAGAGA-3′ | [24] |
| Psuppol-F | 5′-ATAAACCAGCCAGCCGGAA-3′ | This study |
| Psuppol-R | 5′-TTCTGACAACGATCGGAGGA-3′ | This study |
| BE5-F | 5′-TCATGCAGGTGAATGTCGAT-3′ | This study |
| BE5-R | 5′-CTATCCGCAGGAGTTGAACG-3′ | This study |
| BE53-F | 5′-AGATTGATGTTGCCGAGGAC-3′ | This study |
| BE53-R | 5′-TGAAAAAGCTCCGTGAGGTC -3′ | This study |
| BE85-F | 5′-GCGCGGATATTGACATTGAT-3′ | This study |
| BE85-R | 5′-AGGCCGTCGATCTCTATCAC-3′ | This study |
| BE103-F | 5′-ACAAGAAGATGTCGGCCAAG-3′ | This study |
| BE103-R | 5′-TGCTCGCAGAATACAACTGC-3′ | This study |
| BE168-F | 5′-TCGAAAGCGCACTAGATTGA-3′ | This study |
| BE168-R | 5′-AGCCGTAAATATCGGACAGC-3′ | This study |
| InnBXbaI-intF | 5′-CGGTGGCGGCGGCCGCTCTAGAAAAATGCGCAACTGGAAGAT-3′ | This study |
| InnBEcoRI-intR | 5′-CGATAAGCTTGATATCGAATTCATCTGCTCACCAAGCCAATC-3′ | This study |
a Polr: Polymyxin resistant; Kmr: Kanamycin resistant; Smr: Streptomycin resistant; Spr: Spectinomycin resistant; Tcr: Tetracycline resistant; Apr: Ampicillin resistant; inn: incompatible nodulation. b United States Department of Agriculture, Beltsville, MD, USA.