Cloning and expression analysis of an F-box protein gene DoSKP2A in Dendrobium officinale
10.7501/j.issn.0253-2670.2019.08.028
- Author:
Huan LI
1
Author Information
1. Shaanxi Qinling Application Development and Engineering Center of Chinese Herbal Medicine, College of Pharmacy, Shaanxi University of Chinese Medicine
- Publication Type:Journal Article
- Keywords:
Dendrobium officinale Kimura et Migo;
Gene cloning;
QRT-PCR;
Sequence analysis;
SKP2A
- From:
Chinese Traditional and Herbal Drugs
2019;50(8):1960-1965
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To clone an F-box protein namely S phase kinase-associated protein gene DoSKP2A with full length cDNA, in a rare endangered medicinal orchid species Dendrobium officinale, followed by bioinformatics analysis and expression pattern analysis. Methods: RACE technology were used for gene identification. Characteristics of physiochemical properties, conserved domains and subcellular localization of the protein were determined using a series of bioinformatics tools. The analyses of multiple alignment and phylogenetic tree were performed using DNASTAR 7.0 and MEGA 7.0, respectively. Real time quantitative PCR was used for gene expression analysis. Results: DoSKP2A gene was cloned (GenBank accession KU160472). The full length cDNA of DoSKP2A was 1 507 bp in length, and ORF was 1 101 bp, encoding a 366-aa protein with a molecular weight of 39 590 and an isoelectric point of 7.9. The deduced DoSKP2A protein, without transmembrane or signal peptide residues, contained an F-box core domain (26-88), a leucine-rich repeat (202-226), and multiple conserved motifs. DoSKP2A had high identities (64.6%-72.4%) with SKP2As proteins from various plants. DoSKP2A belonged to the monocotyledons subgroup of the SKP2As evolutionary tree. DoSKP2A gene was differentially expressed in the three included organs. The transcripts were more abundant in the roots, with 6.16 fold, then the stems and the lowest in the leaves. Conclusion: The novel full-length F-box protein gene DoSKP2A was obtained, along with bioinformatics and expression characteristics, which provided molecular basis for the growth and development, signal transduction, and stress resistance of D. officinale.
