TABLE 2.
Strains, plasmids, and vectors used in this study, along with the mutants produced
| Strain, vector, or plasmid | Characteristicsa | Source or reference |
|---|---|---|
| E. coli DH5α | supE44 ΔlacU169 (φ80lacZΔM15) hsdR17 recA1 endA1 gyrA96 thi-1 recA1 | 24 |
| Rhizobium sp. strains | ||
| NGR234 | Rifr derivative of Rhizobium sp. strain NGR234, isolated from Lablab purpureus | 53 |
| NGRΩfixF | fixF mutant obtained by inserting an omega interposon into the unique EcoRI site of fixF (Rifr Spr) | 48 |
| NGRΩgM | y4gM mutant obtained by inserting an omega interposon into the unique EcoRI site of y4gM (Rifr Spr) | This work |
| NGRΩhA | y4hA mutant obtained by inserting an omega interposon into the unique BglII site of y4hA (Rifr Spr) | This work |
| NGRΩnodD2 | nodD2 mutant obtained by inserting an omega interposon into the unique BamHI site of nodD2 (Rifr Kmr) | 13 |
| NGRΔnoeE | Deletion mutant of noeE of NGR234 (Rifr Spr) | 25 |
| NGRΔrmlB-wbgA | rmlB-wbgA mutant obtained by deleting a 3,732-bp EcoRV fragment of pXB285 and replacing it with an omega interposon (Rifr Kmr) | This work |
| NGRΩrmlC | rmlC mutant obtained by inserting an omega interposon into the unique HindIII site of rmlC (Rifr Spr) | This work |
| NGRsyrM2::uidA | syrM2 mutant obtained by inserting a uidA-aph cassette into the unique SacI site of syrM2 (Rifr Kmr) | 30 |
| NGRΩttsI | ttsI mutant obtained by inserting an omega interposon into the unique ApaI site of ttsI (Rifr Spr) | 56 |
| Vectors and plasmids | ||
| Lorist 2 | 5.6-kb cosmid vector (Kmr) | 21 |
| pBBR1-MCS5 | Broad-host-range cloning vector (Gmr) | 31 |
| pBluescript KS(+) | Phage f1, lacαZ+ (Apr) | Stratagene, La Jolla, CA |
| pBR-MZHBgM | 2.5-kb fragment containing y4gM cloned into the HindIII-BamHI sites of pBBR1-MCS5, Gmr | This work |
| pBSgFGHI | pKS+ derivative carrying a 5.6-kb BamHI-ApaI fragment covering a region from rmlB (formerly y4gF) to wbgA (formerly y4gI) (Apr) | This work |
| pBSgFGHIΩKm | pBSgFGHI derivative in which the 3.7-kb EcoRV internal fragment containing rmB-wbgA was replaced by an ΩKmr interposon (Apr Kmr) | This work |
| pJQΩgFGHI | pJQ200SK derivative containing a 4.1-kb BamHI-ApaI fragment carrying the 5′ end of y4gF (rmlB), an ΩKmr interposon, and the 3′ end of y4gI (wbgA) (Gmr Kmr) | This work |
| pJQ200 SK | pACYC184-derived (p15A) suicide vector (Gmr) | 44 |
| pJQMZ1 | pJQ200 SK derivative carrying a 3.9-kb PstI fragment from pXB285 and an ΩSpr interposon inserted into the BglII site of y4hA (Gmr Spr) | This work |
| pJQy4gLΩSp | pJQ200 SK derivative carrying a 2.5-kb EcoRV-BamHI fragment and an ΩSpr interposon inserted into the HindIII site of rmlC (formerly y4gL) (Gmr Spr) | This work |
| pJQy4gMΩSp | pJQ200 SK derivative carrying a 4.5-kb cassette containing y4gM in which an ΩSpr interposon was inserted, Gmr Spr | This work |
| pMIGB | 1.4-kb fragment (amplified using the primer pair 5′-GCAGTGGTCTGCCGCATTTC-3′ and 5′-TCCAGTCCGGTCTTGGAGAG-3′) containing the region upstream of y4gM, cloned into pMP220 (Tcr) | This work |
| pMP220 | IncP expression vector containing a promoterless lacZ gene (Tcr) | 52 |
| pMP-NB5 | NB5 cloned into pMP220 as a 1.4-kb XbaI-PstI fragment | 30 |
| pMP-NB6 | NB6 cloned into pMP220 as a 1.1-kb XbaI-PstI fragment | 30 |
| pMP-NB7 | Also called pNBnoeE; NB7 cloned as a 1.0-kb XbaI-BglII fragment in pMP220 | 25 |
| pRK2013 | Helper plasmid containing the ColE1 replicon with RK2 tra genes (Nmr Kmr) | 16 |
| pRK7813 | Broad-host-range Inc P1 cosmid (Tcr) | 27 |
| pRK7813E/B | pRK7813 with EcoRI to BamHI deleted in the polylinker (Tcr) | This work |
| pXB285 | Lorist 2 clone from pNGR234a (Kmr) | 40 |
a
Ampr, ampicillin resistance; Gmr, gentamicin resistance; Kmr, kanamycin resistance; Nmr, neomycin resistance; Rifr, rifampin resistance; Spr spectinomycin resistance; Tcr, tetracycline resistance.