TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Relevant characteristic(s)a | Reference or origin |
|---|---|---|
| Strains | ||
| Rhodococcus sp. strain RHA1 | PCB degrader, BPH+ | 30 |
| Rhodococcus sp. strain SDR1 | bphS mutant of strain RHA1, BPH− | This study |
| R. erythropolis IAM1399 (= ATCC 15963) | Wild type, BPH− | IAM culture collectionb |
| Plasmids | ||
| pBluescript II KS+ | Cloning vector, Apr | Stratagene |
| pG1013F | pBluescript II KS+ with 8.8-kb HindIII fragment of RHA1 carrying bphST | This study |
| pBSL | pBluescript II KS+ with new SpeI site in place of KpnI site | This study |
| pBAB14 | pBSL with 1.4-kb ApaI-BglII fragment of RHA1 carrying bphT | This study |
| pBAB62 | 4.8-kb ApaI fragment carrying bphS inserted into pBAB14 | This study |
| pBAB62ΔSacI | 1.0-kb SacI fragment carrying bphT deleted from pBAB62, leaving bphS | This study |
| pBAS52 | SacI site of pBAB62ΔSacI converted to a SpeI site | This study |
| pET21(+) | Expression vector, Apr | Novagen |
| pETT14 | pET21(+) with SpeI fragment of pBAB14 carrying bphT | This study |
| pETST62 | pET21(+) with SpeI fragment of pBAB62 carrying bphST | This study |
| pKLA1 | Promoter probe vector containing luciferase structural gene, luxAB from V. harveyi, Kmr (neo) | 38 |
| pKLAF1 | pKLA1 with 1.4-kb XhoI-BamHI fragment of RHA1 carrying promoter region of bphA1 | This study |
| pKA851 | pKLAF1 with 8.8-kb HindIII fragment of RHA1 carrying bphST | This study |
| pKAD1-12 | pKLAF1 with deletion derivatives of 8.8-kb HindIII fragment of RHA1 | This study |
| pKLAS1 | pKLAF1 with 5.4-kb SacI fragment of RHA1 carrying bphS | This study |
| pKLAST1 | pKLAF1 with 6.2-kb ApaI-BglII fragment of RHA1 carrying bphST | This study |
| pKLASF | pKLA1 with 0.9-kb PstI-ClaI fragment of RHA1 carrying promoter region of bphS; direction of bphS is identical to that of luxAB reporter gene | This study |
| pKLASR | pKLA1 with 0.9-kb PstI-ClaI fragment of RHA1 carrying promoter region of bphS; direction of bphS is opposite that of luxAB reporter gene | This study |
| pUC-KmD | Gene disruption plasmid, Kmr (aphII) | 29 |
| pUKSD | bphS disruption plasmid pUC-KmD with 1.3-kb ClaI fragment carrying bphS internal fragment of RHA1 | This study |
| pFAJ2574 | Rhodococcus-E. coli shuttle vector, Cmr | 5 |
| pFJS1 | pFAJ2574 with 5.2-kb SpeI fragment of pBAS52 carring bphS | This study |
a
BPH+, growth on biphenyl; BPH−, no growth on biphenyl.
b
IAM, Institute of Applied Microbiology.