title: Isolation and characterization of the first putative peroxidase gene from oilseed rape (Brassica napus) which is highly homologous to HRPC.
abstract: A new gene, designated as BnPrx (GenBank Accession No. DQ078754), was isolated from oilseed rape (Brassica napus) by SMART Rapid Amplification of cDNA Ends (RACE). The full-length cDNA is 1307 bp long and contains a 1062 bp open reading frame (ORF), which encodes a 354 amino acid peroxidase precursor, with a 31 aa N-terminal signal peptide and a 15 aa C-terminal propeptide. The putative protein has a molecular weight of 38.86 kDa and a calculated pI of 5.85. BnPrx shares high identity with HRPC (89%). BnPrx possesses all active residues and two Ca(2+) sites present in Horseradish peroxidase isoenzymes C (HRPC) as well as six N-glycosylation sites. The predicted 3-D structure of BnPrx is very similar to that of HRPC. Assisted by genomic walking technology, the genomic DNA of BnPrx was also cloned, consisting of 3 introns and 4 exons. Thirty-two TATA boxes, 18 CAAT boxes and many cis-elements, such as WUN, MeJR, were found in its promoter region. Southern blot analysis indicated that BnPrx belonged to a small gene family. Northern blot analysis revealed that BnPrx was constitutively expressed in all tested tissues, including roots, stems and leaves, with the high expression in leaves and stems. The expression of BnPrx could be induced by methyl jasmonate (MeJA), salicylic acid (SA), cold and H(2)O(2). The cloning and characterizing of BnPrx might not only help us understand the physiological function and molecular evolution of the large peroxidase gene family more comprehensively, but also provide an alternative way of seeking a more effective and economical substitute for HRPC.
authors.affiliation: State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, Morgan-Tan International Center for Life Sciences, Fudan University, Shanghai , 200433, China.
authors.forename: Weisheng
authors.initials: W
authors.lastname: Wu
authors.forename: Jie
authors.initials: J
authors.lastname: Lu
authors.forename: Yamin
authors.initials: Y
authors.lastname: Wei
authors.forename: Jin
authors.initials: J
authors.lastname: Wang
authors.forename: Juan
authors.initials: J
authors.lastname: Lin
authors.forename: Shuwen
authors.initials: S
authors.lastname: Cao
authors.forename: Xiaofen
authors.initials: X
authors.lastname: Sun
authors.forename: Kexuan
authors.initials: K
authors.lastname: Tang
mesh_terms: D000085:Acetates / Q000378:metabolism; D000595:Amino Acid Sequence; D001483:Base Sequence; D029688:Brassica napus / Q000201:enzymology / Q000235:genetics; D003080:Cold Temperature; D003517:Cyclopentanes / Q000378:metabolism; D018076:DNA, Complementary / Q000302:isolation & purification*; D019143:Evolution, Molecular; D018506:Gene Expression Regulation, Plant; D006735:Horseradish Peroxidase / Q000235:genetics* / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000378:metabolism; D007527:Isoenzymes / Q000737:chemistry / Q000145:classification / Q000235:genetics* / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016877:Oxidants / Q000378:metabolism; D054883:Oxylipins; D009195:Peroxidase / Q000737:chemistry / Q000145:classification / Q000235:genetics* / Q000378:metabolism; D010802:Phylogeny; D010937:Plant Growth Regulators / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011487:Protein Conformation; D020156:Salicylic Acid / Q000378:metabolism; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D014018:Tissue Distribution
chemical_list: D000085:Acetates; D003517:Cyclopentanes; D018076:DNA, Complementary; D007527:Isoenzymes; D016877:Oxidants; D054883:Oxylipins; D010937:Plant Growth Regulators; C072239:methyl jasmonate; D006861:Hydrogen Peroxide; D006735:Horseradish Peroxidase; D009195:Peroxidase; D020156:Salicylic Acid