title: Analyses of glutathione reductase hypomorphic mice indicate a genetic knockout.
abstract: A strain of mice (Gr1a1Neu) that exhibited tissue glutathione reductase (GR) activities that were substantially lower (less than 10% in liver) than the corresponding activities in control mice has been reported. The present report describes characterization of the mutation(s) in the GR gene of these mice. RT-PCR of mRNA from the Neu mice indicated a substantial deletion in the normal GR coding sequence. Southern blots revealed that the deletion involved a region spanning from intron 1 through intron 5. The exact breakpoints of the deletion were characterized by PCR and sequencing through the region encompassing the deletion. The deletion involves nucleotides 10840 through 23627 of the genomic GR gene and functionally deletes exons 2 through 5. In addition, the deletion produces a frame shift in exon 6 and introduces a stop codon in exon 7 that would prevent translation of the remainder of the protein. Consequently, the Neu mice are incapable of producing a functional GR protein and appear to be genetic knockouts for GR. The Neu mice offer live animal models with which to test hypotheses regarding oxidant mechanisms of tissue injury in vivo.
authors.affiliation: Center for Developmental Pharmacology and Toxicology, Children's Research Institute, Columbus, Ohio 43205, USA. rogersl@chi.osu.edu
authors.forename: Lynette K
authors.initials: LK
authors.lastname: Rogers
authors.forename: Toshiya
authors.initials: T
authors.lastname: Tamura
authors.forename: Bryan J
authors.initials: BJ
authors.lastname: Rogers
authors.forename: Stephen E
authors.initials: SE
authors.lastname: Welty
authors.forename: Thomas N
authors.initials: TN
authors.lastname: Hansen
authors.forename: Charles V
authors.initials: CV
authors.lastname: Smith
mesh_terms: D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D015153:Blotting, Western; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005980:Glutathione Reductase / Q000096:biosynthesis / Q000235:genetics*; D020287:In Situ Nick-End Labeling; D007438:Introns / Q000235:genetics; D008099:Liver / Q000201:enzymology / Q000378:metabolism; D051379:Mice; D008809:Mice, Inbred C3H; D018345:Mice, Knockout; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D012333:RNA, Messenger; D004247:DNA; D005980:Glutathione Reductase