A ColE1-compatible expression vector for the production of His-tagged fusion proteins.
Plasmid pDB31 is a ColE1-compatible expression vector based on the p15A origin of replication. It is designed to express His-tagged fusion proteins in cells co-hosting a compatible expression vector. It was constructed by assembling the operator/promoter region plus the 6xHis and the multiple cloning site of pQE31 (QIAGEN) with the p15A origin of replication plus KanR of pGP1-2. The plasmid was found to be stable in Escherichia coli strains BL21 and DH11S. It was used to produce and purify the ferredoxin reductase component of Comamonas testosteroni B-356 biphenyl dioxygenase inside a clone hosting the remaining dioxygenase genes on a compatible plasmid.
Barriault
D
D
INRS-Santé, Université du Québec, Pointe-Claire, Canada.
Sylvestre
M
M
D004274:DNA, Recombinant / Q000235:genetics; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D005287:Ferredoxin-NADP Reductase / Q000096:biosynthesis* / Q000235:genetics; D005822:Genetic Vectors / Q000138:chemical synthesis* / Q000235:genetics; D016946:Gram-Negative Aerobic Rods and Cocci / Q000201:enzymology; D006639:Histidine / Q000235:genetics*; D007506:Iron-Sulfur Proteins; D010105:Oxygenases / Q000096:biosynthesis* / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics
D004274:DNA, Recombinant; D007506:Iron-Sulfur Proteins; D011993:Recombinant Fusion Proteins; D006639:Histidine; D010105:Oxygenases; C074161:biphenyl-2,3-dioxygenase; D005287:Ferredoxin-NADP Reductase