Ferrochelatase Structural Mutant ( Fech m1Pas) in the House Mouse

• Published in: Genomics (San Diego, Calif.) Vol. 16; no. 3; pp. 645 - 648
• Main Authors: Boulechfar, Samia, Lamoril, Jérôme, Montagutelli, Xavier, Guenet, Jean-Louis, Deybach, Jean-Charles, Nordmann, Yves, Dailey, Harry, Grandchamp, Bernard, de Verneuil, Hubert
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.06.1993
• Subjects: Animals; Base Sequence; Cloning, Molecular; DNA; Escherichia coli; Ferrochelatase - genetics; Ferrochelatase - metabolism; Mice; Mice, Inbred BALB C; Molecular Sequence Data; Mutation; Polymerase Chain Reaction; Protoporphyria, Erythropoietic
• ISSN: 0888-7543; 1089-8646
• DOI: 10.1006/geno.1993.1242

The molecular basis of an inherited defect of ferrochelatase in mouse ( Fech m1Pas/Fech m1Pas , described by Tutois et al., 1991, J. Clin. Invest. 88: 1730-1736) was investigated. cDNA clones encoding ferrochelatase, isolated by amplification of the mRNA from the liver of a mutant mouse using the polymerase chain reaction, were sequenced by the dideoxynucleotide chain-termination method. All the clones carried a T to A transversion at nucleotide 293, leading to a methionine to lysine substitution at position 98 in the protein (mutation M98K). Hybridization with allele-specific oligonucleotides (ASOs) confirmed the mutation at the cDNA and genomic levels. Finally, expression of the mutant ferrochelatase protein in E. coli demonstrated a marked deficiency in activity in agreement with the activity of the deficient enzyme in vivo. This Fech m1Pas/Fech m1Pas mutant mouse represents a useful model for studying the pathophysiological feature of the human disease and the first accessible model for gene therapy in the field of porphyrias.