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Characterization of an A-type cyclin-dependent kinase gene from Dendrobium candidum

  • Section Cellular and Molecular Biology
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Abstract

Cyclin-dependent kinases (CDKs) play an essential role in cell cycle regulation during the embryonic and postembryonic development of organisms. To better understand the molecular mechanisms of CDKs involved in embryogenesis regulation in the endangered medicinal plant Dendrobium candidum Wall. ex Lindl., a 1229-bp full-length cDNA of an A-type CDK gene, Denca;CDKA;1, was identified using 3′ rapid amplification of cDNA end (RACE) PCR. Denca;CDKA;1 was predicted to encode a 294 amino acid residue-long protein of 33.76 kDa with an isoelectric point of 7.72. The deduced Denca;CDKA;1 protein contained a conserved serine/threonine-protein kinase domain (S-TKc) and a canonical cyclinbinding “PSTAIRE” motif. Multiple sequence alignment indicated that members of CDKA family from various plants exhibited a high degree of sequence identity ranging from 82% to 93%. A neighbor-joining phylogenetic tree showed that Denca;CDKA;1 was clustered into the plant group and was distant from the animal and fungal groups. The modeled three-dimensional structure of Denca;CDKA;1 exhibited the similar functional structure of a fold consisting of β-sheets and α-helices joined by discontinuous random coils forming two relatively independent lobes. Quantitative real-time PCR analysis revealed that Denca;CDKA;1 transcripts were the most abundant in protocorm-like bodies with 4.76 fold, followed by that in roots (4.19 fold), seeds (2.57 fold), and stems (1.57 fold). This study characterized the novel Denca;CDKA;1 gene from D. candidum for the first time and the results will be useful for further functional determination of the gene.

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Abbreviations

CAK:

CDK-activating kinase

cdc:

cell division cycle

CDK:

cyclin-dependent kinase

CKL:

cyclin dependent kinase like

Ct:

cycle threshold

CTAB:

cetyltrimethyl ammonium bromide

GSP:

gene specific primer

ICK:

inhibitor of CDK

M-MLV RT:

moloney leukemia virus reverse transcriptase

ORF:

open reading frame

PDB:

protein data bank

PLB:

protocorm-like body

qRT-PCR:

quantitative real-time polymerase chain reaction

RACE:

rapid amplification of cDNA ends

S-TKc:

serine/threonine-protein kinase domain

UTR:

un-translated region

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Authors and Affiliations

  1. Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, 100193, People’s Republic of China

    Gang Zhang, Chao Song, Ming-Ming Zhao, Biao Li & Shun-Xing Guo

  2. College of Pharmacy and Shaanxi Provincial Key Laboratory for Chinese Medicine Basis & New Drugs Research, Shaanxi University of Chinese Medicine, Xi’an, Shaanxi, 712046, People’s Republic of China

    Gang Zhang

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  1. Gang Zhang
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  2. Chao Song
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Correspondence to Shun-Xing Guo.

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Zhang, G., Song, C., Zhao, MM. et al. Characterization of an A-type cyclin-dependent kinase gene from Dendrobium candidum . Biologia 67, 360–368 (2012). https://doi.org/10.2478/s11756-012-0016-y

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