Table 2.

Strains and plasmids used in this study

Strain or plasmidRelevant characteristicsaSource or reference
E. coli strains
 DH5αΔ(lacZYA-argF)U169 hsdR17 relA1 supE44 endA1 recA1 thi-1 gyrA96, φ80dlacZΔM15 Life Technologies, Gaithersburg, Md.
 JM109 endA1 recA1 gyrA96 thi hsdR17 relA1 supE44Δ(lac-proAB) mcrA [F′, traD36 proAB+lacI q ZΔM15] 60
 JM109(DE3) endA1 recA1 gyrA96 thi hsdR17 relA1 supE44 Δ(lac-proAB) mcrA [F′,traD36, proAB +,lacI q ZΔM15], λ(DE3)Promega Corp., Madison, Wis.
Plasmids
 pACYC184Cmr Tcr, p15A origin 8
 pT7-5Apr, ColE1 origin, T7 promoter 57
 pT7-7Apr, ColE1 origin, T7 promoter, ribosome-binding site 2
 pUC13Apr, ColE1 origin, lacpromoter 37
 pUC18Apr, ColE1 origin, lac promoter 60
 pUCBM21Apr, ColE1 origin, lacpromoterBoehringer Mannheim Corp.
 pREP1Cmr, p15A origin, T7 promoter and MCS from pT7-5This study
 pDTG124Apr, nahAd under control of the T7 promoter of pT7-7 54
 pDTG141Apr, nahAaAbAcAd under control of the T7 promoter of pT7-5 52
 pDTG149Apr, nahAaAbAc under control of the T7 promoter of pT7-5This study
 pDTG162Cmr, nahAaAbAc under control of the T7 promoter of pREP1This study
 pDTG601AApr, todC1C2BA in pKK223-3 62
 pDTG613Apr,todC2 in pKK223-3 62
 pDTG629Apr, todC2 in pUCBM21This study
 pDTG630Apr, todC2 under control of the T7 promoter of pT7-7This study
 pDTG800Apr, ntdAaAbAcAd in pUC18 39
 pDTG824Apr,ntdAd in pUC18This study
 pDTG850Apr, ntdAaAbAcAd in pUC13This study
 pDTG951Apr, dntAdunder control of the T7 promoter of pT7-6This study
 pDTG953Cmr, dntAaAbAc under control of the T7 promoter of pREP1This study
 pJS48Apr,dntAaAbAcAd under control of the T7 promoter of pT7-6 55
  • a Apr, ampicillin resistance; Cmr, chloramphenicol resistance; Tcr, tetracycline resistance. The nah genes encode NDO fromPseudomonas sp. strain NCIB 9816-4, the ntd genes encode 2NTDO from Pseudomonas sp. strain JS42, and thednt genes encode DNTDO from Burkholderia sp. strain DNT. In each set of genes, Aa encodes the reductase,Ab encodes the ferredoxin, Ac encodes the oxygenase α subunit, and Ad encodes the oxygenase β subunit. The tod genes encode toluene dioxygenase fromP. putida F1. todA encodes the reductase,todB encodes the ferredoxin, todC1 encodes the oxygenase α subunit, and todC2 encodes the oxygenase β subunit.