TABLE 1.
Bacterial strains and plasmids
| Strain or plasmid | Relevant propertiesa | Source or reference |
|---|---|---|
| E. coli strains | ||
| DH5α | F− λ−recA1 Δ(lacZYA-argF)U169 hsdR17 thi-1 gyrA96 supE44 endA1 relA1 φ80dlacZΔM15 | Gibco-BRL |
| S17-1 | thi pro hdsR hdsM+ recA; chromosomal insertion of RP4-2 (Tc::Mu Km::Tn7) | 38 |
| R. palustris strains | ||
| CGA009 | Wild-type strain; spontaneous Cmr derivative | 21 |
| CGA702 | badH::Kmr (polar) | This study |
| CGA720 | badH::Kmr (nonpolar) | This study |
| Plasmids | ||
| pUC18 | High-copy-number cloning vector; ColE1 lacZ Apr | 45 |
| pJQ200KS | Mobilizable suicide vector; sacB Gmr | 35 |
| pUC18K2 | pUC18 containing nonpolar Kmr cassette | 29 |
| pTrcHisA | Six-histidine fusion protein expression vector; ColE1 lacIq Apr | Invitrogen |
| pPE304 | pHC79; cosmid cloning vector containing 35-kb insert including badH; Apr | 10 |
| pDP101 | pUC18 with 2.6-kb fragment from pPE304 containing badH; Apr | This study |
| pDP203 | pDP101 with Kmr gene at the StuI site in badH | This study |
| pDP204 | pJQ200KS; sacB suicide vector containing Kmr cassette at the StuI site of badH | This study |
| pDP212 | pDP101 with nonpolar Kmr gene at the StuI site in badH | This study |
| pDP213 | pJQ200KS; sacB suicide vector with nonpolar Kmr cassette at the StuI site of badH | This study |
| pDPHisH1 | pTrcHisA vector containing badH; Apr | This study |
a
Apr, ampicillin resistance; Gmr, gentamicin resistance; Cmr, chloramphenicol resistance.