Serveur d'exploration sur le phanerochaete

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Identification of a novel cytochrome P-450 gene from the white rot fungus Phanerochaete chrysosporium.

Identifieur interne : 000C08 ( Main/Exploration ); précédent : 000C07; suivant : 000C09

Identification of a novel cytochrome P-450 gene from the white rot fungus Phanerochaete chrysosporium.

Auteurs : S W Kullman [États-Unis] ; F. Matsumura

Source :

RBID : pubmed:9212420

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English descriptors

Abstract

A gene fragment belonging to the cytochrome P-450 superfamily has been cloned and identified from stationary cultures of the filamentous fungus Phanerochaete chrysosporium by reverse transcriptase (RT)-PCR. A set of degenerate primers homologous to highly conserved regions of known cytochrome P-450 sequences were used for initial RT-PCRs. Individual PCR products were cloned, sequenced, and identified as those belonging to the cytochrome P-450 superfamily based on amino acid sequence homologies and the presence of the highly conserved heme binding region. The nucleotide sequence of a single cDNA clone indicated the presence of an open reading frame encoding a partial cytochrome P-450 protein of 208 amino acids. Comparisons of the deduced amino acid sequence of the partial protein to other known cytochrome P-450 sequences indicate that it is the first member of a new family of cytochrome P-450s, designated CYP63-1A. Northern blot analysis suggests that CYP63-1A is expressed under both nitrogen-rich and nitrogen-deficient culture conditions and thus not under the same regulatory constraints as the well-studied lignin and manganese peroxidases. Western blot analyses using antibodies raised to the heme binding region of CYP63-1A indicate that the protein has a molecular mass of approximately 44,000 Da.

DOI: 10.1128/AEM.63.7.2741-2746.1997
PubMed: 9212420
PubMed Central: PMC168569


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Le document en format XML

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<term>Blotting, Western (MeSH)</term>
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<term>Cytochrome P-450 Enzyme System (genetics)</term>
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<term>Heme (genetics)</term>
<term>Heme (immunology)</term>
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<term>Nitrogen (metabolism)</term>
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<term>Similitude de séquences d'acides aminés (MeSH)</term>
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<div type="abstract" xml:lang="en">A gene fragment belonging to the cytochrome P-450 superfamily has been cloned and identified from stationary cultures of the filamentous fungus Phanerochaete chrysosporium by reverse transcriptase (RT)-PCR. A set of degenerate primers homologous to highly conserved regions of known cytochrome P-450 sequences were used for initial RT-PCRs. Individual PCR products were cloned, sequenced, and identified as those belonging to the cytochrome P-450 superfamily based on amino acid sequence homologies and the presence of the highly conserved heme binding region. The nucleotide sequence of a single cDNA clone indicated the presence of an open reading frame encoding a partial cytochrome P-450 protein of 208 amino acids. Comparisons of the deduced amino acid sequence of the partial protein to other known cytochrome P-450 sequences indicate that it is the first member of a new family of cytochrome P-450s, designated CYP63-1A. Northern blot analysis suggests that CYP63-1A is expressed under both nitrogen-rich and nitrogen-deficient culture conditions and thus not under the same regulatory constraints as the well-studied lignin and manganese peroxidases. Western blot analyses using antibodies raised to the heme binding region of CYP63-1A indicate that the protein has a molecular mass of approximately 44,000 Da.</div>
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