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
NDOpDTG162pDTG124nahAaAbAcnahAd
NDO-β2NTDOpDTG162pDTG824nahAaAbAcntdAd
NDO-βDNTDOpDTG162pDTG951nahAaAbAcdntAd
NDO-βTDOpDTG162pDTG630nahAaAbActodC2
NDO-β0pDTG162pT7-7nahAaAbAcNone
DNTDOpDTG953pDTG951dntAaAbAcdntAd
DNTDO-β2NTDOpDTG953pDTG824dntAaAbAcntdAd
DNTDO-βNDOpDTG953pDTG124dntAaAbAcnahAd
DNTDO-βTDOpDTG953pDTG630dntAaAbActodC2
DNTDO-β0pDTG953pT7-7dntAaAbAcNone
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.