A reporter gene construct for studying the regulation of manganese peroxidase gene expression.
The orotidylate decarboxylase (ODase) gene (ura1) from Schizophyllum commune was utilized as a reporter for studying Mn regulation of the manganese peroxidase (MnP) gene (mnp) from the lignin-degrading basidiomycete Phanerochaete chrysosporium. A 1,500-bp fragment of the mnp1 promoter was fused upstream of the coding region of the ODase gene in a plasmid (pAMO) containing the S. commune ade5 gene as a selectable marker. pAMO was used to transform a P. chrysosporium ade1 ura11 mutant lacking endogenous ODase activity. When the P. chrysosporium transformant was grown in nitrogen-limited, Mn(II)-sufficient cultures, ODase activity was detected only during secondary metabolic growth and the pattern of ODase expression was similar to that of endogenous MnP. When Mn was added to 6-day-old nitrogen-limited, Mn-deficient cultures, both ODase activity and MnP activity were induced synchronously with maximal activity at 30 h. Growth in high-nitrogen-concentration medium suppressed the induction of both the ODase and endogenous MnP. These results indicate that this promoter-reporter construct can be used to study the regulation of the mnp gene.
Department of Chemistry, Biochemistry, and Molecular Biology, Oregon Graduate Institute of Science and Technology, Portland 97291-1000.
B J
BJ
Godfrey
L
L
Akileswaran
M H
MH
Gold
D001483:Base Sequence; D001487:Basidiomycota / Q000201:enzymology / Q000235:genetics*; D016384:Consensus Sequence; D004790:Enzyme Induction; D005656:Fungal Proteins / Q000096:biosynthesis* / Q000235:genetics; D005800:Genes, Fungal; D005813:Genes, Synthetic; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D009584:Nitrogen / Q000378:metabolism; D009964:Orotidine-5'-Phosphate Decarboxylase / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics; D010544:Peroxidases / Q000096:biosynthesis* / Q000235:genetics; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics; D014170:Transformation, Genetic
D005656:Fungal Proteins; D011993:Recombinant Fusion Proteins; D010544:Peroxidases; C051129:manganese peroxidase; D009964:Orotidine-5'-Phosphate Decarboxylase; D009584:Nitrogen