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Cloning, characterization and localization of a novel basic peroxidase gene from Catharanthus roseus

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Title Cloning, characterization and localization of a novel basic peroxidase gene from Catharanthus roseus
 
Creator Kumar, Santosh
Dutta, Ajaswrata
Sinha, Alok Krishna
Sen, Jayanti
 
Subject Catharanthus roseus
organ specific
peroxidase
terpenoid indole alkaloid
subcellular localization
 
Description Catharanthus roseus (L.) G. Don produces a number of biologically active
terpenoid indole alkaloids via a complex terpenoid indole alkaloid biosynthetic pathway. The final dimerization step of this pathway, leading to the
synthesis of a dimeric alkaloid, vinblastine, was demonstrated to be catalyzed by a basic peroxidase. However, reports of the gene encoding this
enzyme are scarce for C. roseus. We report here for the first time the cloning, characterization and localization of a novel basic peroxidase, CrPrx,
from C. roseus. A 394 bp partial peroxidase cDNA (CrInt1) was initially
amplified from the internodal stem tissue, using degenerate oligonucleotide
1 primers, and cloned. The full-length coding region of CrPrx cDNA was
isolated by screening a leaf-specific cDNA library with CrInt1 as probe.
The CrPrx nucleotide sequence encodes a deduced translation product of
330 amino acids with a 21 amino acid signal peptide, suggesting that CrPrx
is secretory in nature. The molecular mass of this unprocessed and unmodified deduced protein is estimated to be 37.43 kDa, and the pI value is 8.68.
CrPrx was found to belong to a ‘three intron’ category of gene that
encodes a class III basic secretory peroxidase. CrPrx protein and mRNA
were found to be present in specific organs and were regulated by different
stress treatments. Using a beta-glucuronidase–green fluorescent protein fusion
of CrPrx protein, we demonstrated that the fused protein is localized in
leaf epidermal and guard cell walls of transiently transformed tobacco. We
propose that CrPrx is involved in cell wall synthesis, and also that the gene
is induced under methyl jasmonate treatment. Its potential involvement in
the terpenoid indole alkaloid biosynthetic pathway is discussed.
We thank the
Department of Biotechnology (DBT), Government of
India for its financial support.
 
Date 2013-11-06T04:15:20Z
2013-11-06T04:15:20Z
2007
3 Januay 2007
 
Type Article
 
Identifier FEBS Journal, 274: 1290-1303
http://hdl.handle.net/123456789/73
 
Language en
 
Publisher Wiley-Blackwell