TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Relevant characteristicsa | Reference or source |
|---|---|---|
| Strains | ||
| E. coli | ||
| DH5α | F− φ80DlacZΔM15 endA1 recA1 hsdR17(rK− mK−) supE44 thi-1 gyrA96 Δ(lacZYA-argF) | 49 |
| HB101 | F−hsd20
(rK− mK−)
supE44 ara-14 galK2 lacY1 proA2 rpsL20 Smrxyl-5 leuB6 mtl-1 lambda−recA13 thy thi |
49 |
| FAJ1323 | DH5α containing pFAJ1323; Apr | This study |
| A. tumefaciens NT1(pJM749, pSVB33) | Indicator strain for detecting AHLs; Kmr Cbr | 44 |
| R. leguminosarum bv. viciae | ||
| 248 | Sensitive to bacteriocin small; Rifr | |
| RBL1309 | small producing | 57 |
| RBL1376 | RBL1309 Tn5 mutant; small | 57 |
| R. etli | ||
| CNPAF512 | Wild-type isolate; Nalr | Embrapa, Brazil |
| FAJ1328 | raiI::gusA-Km mutant, convergently oriented; Nalr Kmr | This study |
| FAJ1329 | raiR mutant with gusA-Km in reverse orientation; Nalr Kmr | This study |
| CNPAF512::pFAJ1328 | cis merodiploid raiI::gusA-Km; Gmr Nalr Kmr | This study |
| CNPAF512::pFAJ1334 | cis merodiploid ORF1::gusA-Km; two copies of raiI/raiR; Gmr Nalr Kmr | This study |
| Plasmids | ||
| pHV200 | 8.8-kb SalI fragment with V. fischeri ES184 lux regulon; hybridization probe | 25 |
| pUC18 | ColE1 origin; cloning vehicle; Apr | 62 |
| pLAFR1 | IncP1 cos oriT; Tcr | 17 |
| pCR™2.1 TA | ColE1 origin, F1 origin; cloning vehicle for PCR products; Apr Kmr | Invitrogen |
| pWM6 | Source of gusA-Kmr cassette; Apr Kmr | 37 |
| pJQ200uc1 | Suicide vector, sacB; Kmr Gmr | 47 |
| pRK2013 | ColE1 replicon with RK2 transfer genes; Kmr | 15 |
| pFAJ1322 | Cosmid clone, pLAFR1 derivative, AHL+; Tcr | This study |
| pFAJ1323 | SalI fragment from pFAJ1322 in pUC18; AHL+; Apr | This study |
| pFAJ1326 | PCR fragment (raiI raiR ORF1) in pCR2.1 TA; Apr | This study |
| pFAJ1327 | PCR fragment (raiI raiR ORF1) in pJQ200uc1 via NotI; Gmr | This study |
| pFAJ1328 | pFAJ1327; raiI::gusA-Kmr in XhoI; Kmr Gmr | This study |
| pFAJ1329 | pFAJ1327; raiR with gusA-Kmr insertion in NdeI; Kmr Gmr | This study |
| pFAJ1334 | pFAJ1327; ORF1::gusA-Kmr in BamHI; Kmr Gmr | This study |
a
Nalr, nalidixic acid resistant; Apr, ampicillin resistant; Kmr, kanamycin-neomycin resistant; Tcr, tetracycline resistant; Gmr, gentamicin resistant; Cbr, carbenicillin resistant; Rifr, rifampin resistant.