Isolation and expression analysis of caffeoyl CoA O-methyltransferase gene in Dendrobium officinale
10.7501/j.issn.0253-2670.2014.08.019
- Author:
Gang ZHANG
1
Author Information
1. Shaanxi Provincial Key Laboratory for Chinese Medicine Basis and New Drugs Research, College of Pharmacy, Shaanxi University of Chinese Medicine
- Publication Type:Journal Article
- Keywords:
Caffeoyl-CoA O-methyltransferase;
Dendrobium officinale Kimura et Migo;
Gene cloning;
Real time quantitative PCR;
Sequence analysis
- From:
Chinese Traditional and Herbal Drugs
2014;45(8):1143-1149
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To isolate the caffeoyl CoA O-methyltransferase (CCoAOMT) gene (DoOMT) in a rare endangered medicinal orchid species Dendrobium officinale, and to carry out the bioinformatics and expression mode analysis. Methods: RT-PCR and RACE technologies were used to obtain the full length cDNA of DoOMT gene. The characteristics of physiochemical properties, conserved domains, and three dimensional structure of the deduced DoOMT protein were determined using a series of bioinformatic tools. The analyses of multiple alignment and phylogenetic tree were performed using DNASTAR 6.0 and MEGA 4.0 softwares, respectively. Real time quantitative PCR was used for gene expression analysis. Results: The full length cDNA of DoOMT was 1 005 bp in length and encoded a 239-amino acid protein with a molecular weight of 2.708×104 and an isoelectric point of 5.03; The deduced DoOMT protein, without transmembrane or signal peptide residues, contained the oxygen methyltransferase family 3, methyltransferase conserved domains (13-238 and 31-238), and eight conserved signature sequences. DoOMT had high identities (49.4%-78.7%) with CCoAOMTs proteins from various plants; DoOMT belonged to the 1b subgroup of the CCoAOMT evolutionary tree, and was closely related to the monocot Vanilla planifolia. DoOMT transcripts were constitutively expressed in the leaves, stems, and roots. The transcription level of DoOMT was markedly higher than that in the leaves with 4.562 fold, whereas the transcript amount showed no significant difference in the leaves and roots. Conclusion: Molecular characterization of DoOMT will be useful for the further functional determination of the gene involving in the secondary metabolism along with growth and development in D. officinale.
