title: Assembly and functional expression of murine glutathione reductase cDNA: a sequence missing in expressed sequence tag libraries.
abstract: Glutathione reductase (GR) is a chemotherapeutic target. Murine GRcDNA, which contains 85% GC in the 38 codons following the start codon, was assembled from the PCR-amplified exon 1 and a downstream cDNA prior to expression in Escherichia coli as a His(6)-tagged protein. Recombinant GR, an FAD-containing homodimer, corresponds in its enzymic and spectral properties to GR isolated from murine Ehrlich ascites tumor cells. Another cDNA, representing GR with a mitochondrial targeting sequence, yielded two distinct enzymically active expression products.
authors.affiliation: Center of Biochemistry, Heidelberg University, Im Neuenheimer Feld 328, D-69120, Heidelberg, Germany.
authors.forename: R
authors.initials: R
authors.lastname: Iozef
authors.forename: K
authors.initials: K
authors.lastname: Becker
authors.forename: C C
authors.initials: CC
authors.lastname: Boehme
authors.forename: R H
authors.initials: RH
authors.lastname: Schirmer
authors.forename: D
authors.initials: D
authors.lastname: Werner
mesh_terms: D000818:Animals; D002286:Carcinoma, Ehrlich Tumor / Q000201:enzymology / Q000235:genetics / Q000473:pathology; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D020224:Expressed Sequence Tags; D020862:GC Rich Sequence; D015723:Gene Library; D005980:Glutathione Reductase / Q000096:biosynthesis* / Q000235:genetics*; D051379:Mice; D008928:Mitochondria / Q000378:metabolism; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D014407:Tumor Cells, Cultured
chemical_list: D018076:DNA, Complementary; D011994:Recombinant Proteins; D005980:Glutathione Reductase