Table 3.

Two-plasmid expression system for wild-type and hybrid dioxygenase production

Dioxygenase designationPlasmidsa present in JM109(DE3)Dioxygenase genes present in:
Plasmid APlasmid BPlasmid APlasmid B
NDOpDTG162pDTG124 nahAaAbAc nahAd
NDO-β2NTDO pDTG162pDTG824 nahAaAbAc ntdAd
NDO-βDNTDO pDTG162pDTG951 nahAaAbAc dntAd
NDO-βTDO pDTG162pDTG630 nahAaAbAc todC2
NDO-β0 pDTG162pT7-7 nahAaAbAc None
DNTDOpDTG953pDTG951 dntAaAbAc dntAd
DNTDO-β2NTDO pDTG953pDTG824 dntAaAbAc ntdAd
DNTDO-βNDO pDTG953pDTG124 dntAaAbAc nahAd
DNTDO-βTDO pDTG953pDTG630 dntAaAbAc todC2
DNTDO-β0 pDTG953pT7-7 dntAaAbAc None
Vector controlpREP1pT7-7NoneNone
  • a In each case, plasmid A is a derivative of pREP1 (Cmr) carrying the genes encoding reductase, ferredoxin, and oxygenase α subunit and plasmid B is a ColE1 derivative (Apr) carrying the oxygenase β subunit gene. See Table 2 and Materials and Methods for details of plasmid constructions and gene designations.