Cloning and expression analysis of a magnesium transporter gene in Dendrobium officinale
10.7501/j.issn.0253-2670.2014.23.017
- Author:
Gang ZHANG
1
Author Information
1. Shaanxi Provincial Key Laboratory for Chinese Medicine Basis & New Drugs Research College of Pharmacy, Shaanxi University of Chinese Medicine
- Publication Type:Journal Article
- Keywords:
Dendrobium officinale Kimura et Migo;
Gene cloning;
Magnesiumion transporter;
Real time quantitative PCR;
Sequence analysis
- From:
Chinese Traditional and Herbal Drugs
2014;45(23):3443-3448
- CountryChina
- Language:Chinese
-
Abstract:
Objective: This study was carried out to clone a magnesium transporter gene DoMGT1 from a medicinally important endangered orchid species Dendrobium officinale, followed by bioinformatics and expression analysis. Methods: RT-PCR and RACE technologies were used to isolate the full-length gene. The characteristics of physicochemical properties, conserved domains, and subcellular localization of DoMGT1 protein were determined using a series of bioinformatics 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 employed for gene expression analysis. Results: The full-length cDNA of DoMGT1 was 1 625 bp in size, and encoded a 425-aa peptide chain with a molecular weight of 47 400 and an isoelectric point (pI) of 4.79; The deduced DoMGT1 protein contained the magnesiumion transporter CorA-like (313-420) and MRS2/LEP10 (27-420) conserved domain. DoMGT1 was highly similar (46%-63%) to MGTs genes from various plants. The deduced DoMGT1 protein was closely related to rice OsMGTD, OsMGTE, and OsMGTF proteins, and belonged to one clade of plant MGTs gene together with AtMGT4. DoMGT1 transcripts were constitutively expressed in leaves, stems, and roots of D. officinale. The transcription level of DoMGT1 in leaves was the highest (3.46 fold higher than that of roots), followed by that of stems (1.54 fold). Conclusion: The full-length DoMGT1 gene has been successfully cloned. The high expression level of DoMGT1 in D. officinale leaves suggests that the gene might play a vital regulatory role in the leaves.
