Molecular Characterization of A Protein Phosphatase 2C Gene of Subgroup A in Dendrobium officinale / 中国药学杂志

ABSTRACT

OBJECTIVE: To characterize a protein phosphatase (PP) encoding gene DoPP2C2 in a rare endangered medicinal orchid species Dendrobium officinale, followed by bioinformatics analysis and expression profiling. METHODSE: RACE and RT-PCR were used to isolate the full length cDNA of DoPP2C2. The physiochemical properties, conserved domains and subcellular localization of the deduced DoPP2C2 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 6.0 softwares, respectively. Quantitative PCR was used for gene expression analysis. RESULTS: The full length cDNA of DoPP2C2 (GenBank accession KT957553) was 1 624 bp in length, and encoded a 387-aa protein with a molecular weight of 42.76×103 and an isoelectric point of 7.03. The deduced DoPP2C2 protein sequence had two PP2C domains (91-152, 216-376), which are all conserved among the PP2Cs. The protein without a signal peptide or a transmembrane region, was predicted to locate in nucleus with hydropathicity. DoPP2C2 had high identities (47.8%-73.4%) with various PP2C proteins from several plants. DoPP2C2 protein belonged to the subgroup A of Arabidopsis and rice PP2C evolutionary tree, and was closely related to OsPP2C30; DoPP2C2 was differentially expressed in the three included organs. The transcripts were less in the roots and stems, being 0.76 and 0.59 folds, respectively, of that in the leaves. CONCLUSION: The molecular characteristics of a subgroup protein phosphatase encoding gene DoPP2C2 of the full length cDNA in D. officinale was obtained. The results will be useful for further functional determination of the gene involving in the growth and development of D. officinale.