Cloning and expression analysis of a zinc-regulated transporters (ZRT), iron-regulated transporter (IRT)-like protein encoding gene in Dendrobium officinale

ZHANG Gang1,2 LI Yi-min1 LI Biao2 ZHANG Da-wei2 GUO Shun-xing2

(1.College of Pharmacy and Shaanxi Provincial Key Laboratory for Chinese Medicine Basis & New Drugs Research, Shaanxi University of Chinese Medicine, Xi'an, China 712046)
(2.Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences-Peking Union Medical College, Beijing, China 100193)

【Abstract】The zinc-regulated transporters (ZRT), iron-regulated transporter (IRT)-like protein (ZIP) plays an important role in the growth and development of plant. In this study, a full length cDNA of ZIP encoding gene, designed as DoZIP1 (Gen Bank accession KJ946203), was identified from Dendrobium officinale using RT-PCR and RACE. Bioinformatics analysis showed that DoZIP1 consisted of a 1 056 bp open reading frame (ORF) encoded a 351-aa protein with a molecular weight of 37.57 kDa and an isoelectric point (pI) of 6.09. The deduced DoZIP1 protein contained the conserved ZIP domain, and its secondary structure was composed of 50.71% alpha helix, 11.11% extended strand, 36.18% random coil, and beta turn 1.99%. DoZIP1 protein exhibited a signal peptide and eight transmembrane domains, presumably locating in cell membrane. The amino acid sequence had high homology with ZIP proteins from Arabidopsis, alfalfa and rice. A phylogenetic tree analysis demonstrated that DoZIP1 was closely related to AtZIP10 and OsZIP3, and they were clustered into one clade. Real time quantitative PCR analysis demonstrated that the transcription level of DoZIP1 in D. officinale roots was the highest (4.19 fold higher than that of stems), followed by that of leaves (1.12 fold). Molecular characters of DoZIP1 will be useful for further functional determination of the gene involving in the growth and development of D. officinale.

【Keywords】 Dendrobium officinale; zinc-regulated transporters iron-regulated transporter-like protein (ZIP); gene clone; sequence analysis; real time quantitative PCR;

【DOI】

【Funds】 the National Natural Science Foundation of China (Grant Nos. 31070300, 31101608, 31170314) Youth star of science and technology of Shaanxi science and technology program

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This Article

ISSN:1001-5302

CN: 11-2272/R

Vol 40, No. 01, Pages 42-47

January 2015

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Abstract

  • 1 Materials and Methods
  • 2 Results and Analyses
  • 3 Discussion
  • References