Cloning and in silico analysis of a cinnamyl alcohol dehydrogenase gene in Pennisetum purpureum.
Lignin is a major constituent of plant cell walls and indispensable to the normal growth of a plant. However, the presence of lignin complicates the structure of the plant cell walls and negatively influences pulping industry, lignocellulose utilization as well as forage properties. Cinnamyl alcohol dehydrogenase (CAD), a key enzyme involved in lignin biosynthesis, catalyses the last step in monolignol synthesis and has a major role in genetic regulation of lignin production. In the present study, a 1 342-bp cDNA fragment of CAD gene, named PpCAD, was isolated from Pennisetum purpureum using strategies of homologous clone and rapid amplification of cDNA end. It was translated into an intact protein sequence including 366 amino acid residues by ORF Finder. The genomic full-length DNA of PpCAD was a 3 738-bp sequence containing four exons and three introns, among which the 114-bp exon was considered to be a conserved region compared with other CADs. Basic bioinformatic analysis presumed that the PpCAD was a nonsecretory and hydrophobic protein with five possible transmembrane helices. The phylogenetic analysis indicated that the PpCAD belonged to the class of bona fide CADs involved in lignin synthesis and it showed a high similarity (nearly 90%) with CAD protein sequences of Sorghum bicolor, Panicum virgatum and Zea mays in Gramineae. Furthere, PpCAD amino acid sequence was demonstrated to have some conserved motifs such as Zn-binding site, Zn-catalytic centre and NADP(H) binding domain after aligning with other bona fide CADs. Three-dimensional homology modelling of PpCAD showed that the protein had some exclusive features of bona fide CADs.
College of Agriculture, South China Agricultural University, Guangzhou 510642, People's Republic of China. xiexmbs@scau.edu.cn.
Ran
R
Tang
Xiang-Qian
XQ
Zhang
You-Han
YH
Li
Xin-Ming
XM
Xie
D000429:Alcohol Oxidoreductases / Q000235:genetics*; D000595:Amino Acid Sequence; D020134:Catalytic Domain; D003001:Cloning, Molecular; D019295:Computational Biology; D005091:Exons; D023061:Gene Order; D017343:Genes, Plant; D007438:Introns; D008958:Models, Molecular; D008969:Molecular Sequence Data; D005810:Multigene Family; D031782:Pennisetum / Q000235:genetics*; D010802:Phylogeny; D011487:Protein Conformation; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
D000429:Alcohol Oxidoreductases; C018656:cinnamyl alcohol dehydrogenase