High-level expression of human dihydropteridine reductase (EC 1.6.99.7), without N-terminal amino acid protection, in Escherichia coli.
The cDNA coding for human dihydropteridine reductase [Dahl, Hutchinson, McAdam, Wake, Morgan & Cotton (1987) Nucleic Acids Res. 15, 1921-1936] was inserted downstream of tandem bacteriophage lambda PR and PL promoters in Escherichia coli vector pCE30. Since pCE30 also expresses the lambda c1857ts gene, transcription may be controlled by variation of temperature. The recombinant plasmid in an E. coli K12 strain grown at 30 degrees C, then at 45 degrees C, directed the synthesis of dihydropteridine reductase to very high levels. The protein was soluble, at least as active as the authentic human enzyme, and lacked the N-terminal amino acid protection.
Armarego
W L
WL
Division of Biochemistry and Molecular Biology, John Curtin School of Medical Research, Australian National University, Canberra.
Cotton
R G
RG
Dahl
H H
HH
Dixon
N E
NE
D003001:Cloning, Molecular; D004093:Dihydropteridine Reductase / Q000032:analysis*; D004926:Escherichia coli / Q000201:enzymology*; D009247:NADH, NADPH Oxidoreductases / Q000032:analysis*
D004093:Dihydropteridine Reductase; D009247:NADH, NADPH Oxidoreductases