Assembly and functional expression of murine glutathione reductase cDNA: asequence missing in expressed sequence tag libraries

Glutathione reductase (GR) is a chemotherapeutic target. Murine GRcDNA, whi ch contains 85% GC in the 38 codons following the start codon, was assemble d from the PCR-amplified exon 1 and a downstream cDNA prior to expression i n Escherichia coli as a His(6)-tagged protein. Recombinant GR, an FAD-conta ining homodimer, corresponds in its enzymic and spectral properties to GR i solated from murine Ehrlich ascites tumor cells. Another cDNA, representing GR with a mitochondrial targeting sequence, yielded two distinct enzymical ly active expression products. (C) 2000 Elsevier Science B.V. All rights re served.