Scientia Agricultura Sinica ›› 2013, Vol. 46 ›› Issue (9): 1881-1891.doi: 10.3864/j.issn.0578-1752.2013.09.016

• HORTICULTURE • Previous Articles     Next Articles

Cloning and Expression Analysis of Eukaryotic Translation Initiation Factor 4E Gene and Screening of the Interactive Protein from Chrysanthemum×morifolium

 LOU  Wang-Huai, AN  Juan, SONG  Ai-Ping, CHEN  Su-Mei, JIANG  Jia-Fu, CHEN  Fa-Di, FANG  Wei-Min, GUAN  Zhi-Yong   

  1. College of Horticulture, Nanjing Agricultural University, Nanjing 210095
  • Received:2012-04-13 Online:2013-05-01 Published:2013-03-06

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Abstract: 【Objective】In order to study the expression patterns and function of translation initiation factor 4E in the Chrysanthemum, the eukaryotic translation initiation factor 4E gene was cloned from Chrysanthemum× morifolium ‘Jinba’ and its expression was analyzed, and the proteins interacting with the CmeIF4E protein were screened subsequently.【Method】CmeIF4E gene was cloned from Chrysanthemum by RT-PCR and RACE, the primers were designed according to the conserve motifs of eIF4E from several plant species, whose sequence of eIF4E has been reported. The expression patterns of CmeIF4E was analyzed by quantitative real-time PCR and subcellular localization, then screened the interacting proteins via yeast two-hybrid system.【Result】The eIF4E gene cloned from Chrysanthemum is 914 bp in length, ORF 654 bp, encoding 218 amino acids, named CmeIF4E. The sequences were submitted into GenBank database with accession No. JQ904591. The amino acid sequence analysis showed that the genetic relationships between Chrysanthemum and lettuce were the closest, and this result was consistent with plant taxonomy. Quantitative real-time PCR analysis showed that the CmeIF4E gene expressed in all tissues of Chrysanthemum ‘jinba’, the expression level of CmeIF4E in the young roots was the highest, next by the leaves, and was the lowest in stems. The subcellular localization assay showed that CmeIF4E was expressed in the cell nucleus, cytoplasm and cell membrane. The interacting proteins of CmeIF4E were related to the pathways of translation and post-translational modifications, photosynthesis, stress tolerance and defense.【Conclusion】CmeIF4E was constitutively expressed in Chrysanthemum tissues, while transient expression showed that CmeIF4E protein was located in the nucleus, cytoplasm and cell membrane. The candidate proteins screened by Y2H system showed that CmeIF4E might play an important role in translation initiation, photosynthetic system, plant resistance defense. All these results provided an important basis for the further study of the role of the CmeIF4E protein in Chrysanthemum.

Key words: Chrysanthemum ×, morifolium , eIF4E , virus , expression analysis , subcellular localization , yeast two-hybrid system

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