Abstract
A cDNA clone of a manganese peroxidase (MnP) from Phanerochaete chrysosporium was isolated and characterized. The cDNA contains 1314 nucleotides excluding the poly(A) tail and the coding region has 68% G + C content. The deduced mature MnP protein contains 357 amino acids and is preceded by a 21-amino acid leader sequence. The experimentally determined N-terminal sequence of the purified MnP-1 protein, pI = 4.9, corresponds to the deduced N-terminal sequence of the gene. The Mr of the mature MnP-1 deduced from the cDNA is 37,439, which is approximately 81.4% of the experimentally determined molecular weight. The difference is due to glycosylation and a single potential N-glycosylation site with the general sequence Asn-X-Thr/Ser is present in the deduced MnP-1 sequence. Consistent with the peroxidase mechanism of MnP, the proximal histidine, the distal histidine, and the distal arginine are all conserved and regions flanking these residues display homology with other peroxidases. Northern blot analysis indicates that MnP expression is controlled by nutrient nitrogen at the level of transcription. Southern blot hybridization analysis suggests that MnP-1 is a member of a family of MnP genes.
Highlights
AcDNA clone of a manganese peroxidase (MnP) been sequenced [8, 9]
We residues display homology with other peroxidases. report the nucleotide sequence of a cDNA clone for one of the Northern blot analysis indicates that MnP expression MnP isozymesand presentpreliminary information about the is controlled by nutrient nitrogen at the level of tran- regulation of MnP andabout the MnPgene family
All but 162 bp (12%) of the sequence were obtained from both strands of the cDNA; the remainder was sequenced (5 X) without apparent ambiguity
Summary
The second most abundant natural polymer, is a complex optically inactive and random phenylpropanoid matrix that comprises 20-30% of woody plants [1,2]. A restriction map for replicative form DNA from the MnP-1 clone was produced by standard procedures [21, 37] and restriction fragments of the MnP-1cDNA were subcloned directly from gels into mp or mp replicative form. Both orientations of one 260-bp PstI fragment from the MnP-1coding region gave blue rather than clear plaques [37]. An EcoRI insert representing the entire MnP-1 cDNA wascut out of mp replicative form and isolated from an agarose gel by glass powder extraction [44].This DNA wasused as a templatefor randomprimed synthesis of 32P-labeledprobes [45]
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
