TABLE 1.
Strains and plasmids
| Strain or plasmid | Relevant characteristicsa | Reference or source |
|---|---|---|
| Strains | ||
| E. coli | ||
| XL1-Blue | recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 lac[F′ proAB lacIqZΔM15 Tn10 (Tcr)] | StrateGene |
| PIR2 | F− Δlac169 rpoS(Am) robA1 creC510 hsdR514 endA recA1 uidA(ΔMluI)::pir | Invitrogen |
| A. tumefaciens | ||
| A136 | C58 background without Ti plasmid | 63 |
| A348 | C58 background carrying pTiA6 | 63 |
| AB140 | Spontaneous AS-hypersensitive mutant of A348 | 37 |
| AB144 | Spontaneous AS-hypersensitive mutant of A348 | 37 |
| AB147 | Spontaneous AS-hypersensitive mutant of A348 | 37 |
| AB300 | A348 with deletion of chvE | 25 |
| AB3003 | AB300 with gfp ORF inserted between virE2 and virE3 | This study |
| AB3003C | AB3003 with β-lactamase ORF from pBBR1MCS4 inserted after virB1 start codon | This study |
| AB3009 | AB3003C with sacB ORF from pK18mobsacB inserted after virB2 start codon | This study |
| AB3012 | AB3009 with chvE returned to its chromosomal location | This study |
| AB3016 | AB3012 with mexE deletion | This study |
| AB3018 | A348 with mexE deletion | This study |
| MDG165 | 358mx virE::[lacZ::gfp] | 61 |
| Plasmids | ||
| pBBR1MCS2 | Broad-host-range plasmid cloning vector; Kanr | 39 |
| pBBR1MCS4 | Broad-host-range plasmid cloning vector; Ampr | 39 |
| pK18mobsacB | Suicide plasmid vector; Kanr | 62 |
| pYW15C | IncW/ColE1 expression vector with PN25 promoter; Carbr | 43 |
| pRL27 | Tn5-RL27 (Kanr oriR6K) delivery vector | 38 |
| pAB302 | pYW15C with mexE expressed from PN25 promoter; Carbr | This study |
| pAB303 | pK18mobsacB carrying sequences flanking mexE orf; Kanr | This study |
| pJZ150 | chvE plus flanking sequences in pK18mobsacB; Kanr | This study |
| pJZ151 | β-Lactamase ORF fused to virB1 start codon plus flanking upstream and downstream virB1 sequences in pK18mobsacB; Kanr | This study |
| pJZ152 | sacB ORF fused to virB2 start codon plus flanking virB1, virB2, and virB3 sequences in pK18mobsacB; Kanr | This study |
| pJZ153 | gfp ORF (including start codon) plus virE2 and virE3 sequences in pK18mobsacB; Kanr | This study |
a
Carbr, carbenicillin resistance; Kanr, kanamycin resistance.