Assembly and functional expression of murine glutathione reductase cDNA: a sequence missing in expressed sequence tag libraries
Copyright policy )Assembly and functional expression of murine glutathione reductase cDNA: a sequence missing in expressed sequence tag libraries
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.
- Helmholtz Association of German Research Centres Germany
- Heidelberg University Germany
- German Cancer Research Center Germany
Mitochondrial targeting sequence, DNA, Complementary, Molecular Sequence Data, Polymerase Chain Reaction, Chromatography, Affinity, Mice, Escherichia coli, Tumor Cells, Cultured, Animals, Cloning, Molecular, Carcinoma, Ehrlich Tumor, Molecular Biology, Flavin spectra, Gene Library, Expressed Sequence Tags, GC rich sequence, Sequence Homology, Amino Acid, Mouse glutathione reductase, Recombinant Proteins, GC Rich Sequence, Mitochondria, Brain tumor, Glutathione Reductase, Murine malaria, Molecular Medicine
Mitochondrial targeting sequence, DNA, Complementary, Molecular Sequence Data, Polymerase Chain Reaction, Chromatography, Affinity, Mice, Escherichia coli, Tumor Cells, Cultured, Animals, Cloning, Molecular, Carcinoma, Ehrlich Tumor, Molecular Biology, Flavin spectra, Gene Library, Expressed Sequence Tags, GC rich sequence, Sequence Homology, Amino Acid, Mouse glutathione reductase, Recombinant Proteins, GC Rich Sequence, Mitochondria, Brain tumor, Glutathione Reductase, Murine malaria, Molecular Medicine
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