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ACTA HORTICULTURAE SINICA ›› 2012, Vol. 39 ›› Issue (1): 143-150.

• Research Notes • Previous Articles     Next Articles

Cloning and Expression Analysis of PaGAI gene of DELLA Protein from Prunus avium

ZHONG Fei1,SHEN Xin-jie1,LIU Fang1,YUAN Hua-zhao1,LIU Lan-ying2,and LI Tian-hong1,*   

  1. (1College of Agronomy and Biotechnology,Key Laboratory of Stress Physiology and Molecular Biology for Fruit Trees of Beijing,China Agricultural University,Beijing 100193,China;2Laboratory of Plant Tissue Culture Technology,Haidian District,Beijing 100091,China)
  • Received:2011-09-15 Revised:2011-11-24 Online:2012-01-25 Published:2012-01-25
  • Contact: LI Tian-hong

Abstract: DELLA protein is a key negative regulation factor of gibberellins(GAs)signal transduction pathway. A whole cDNA sequence of DELLA protein gene,named PaGAI,was cloned from Prunus avium L. tender leaves using RT-PCR and RACE-PCR. The cDNA of PaGAI had a length of 2 310 bp and contain an opening-reading frame(ORF)of 1 788 bp,which was supposed to encode a 595 amino-acid residues polypeptide of 64 kD. Bioinformatics analysis showed that the protein encoded by PaGAI had a conservative structural domain of DELLA protein. It had two strict conservative amino acid domains DELLA and VHYNP in the N terminal and three repression regions of VHVID,RVER and SAW in the C terminal. PaGAI,with high homologous with the DELLA proteins in some other plants,and showed the highest homology of 84% with Malus domestica. The prokaryotic expression system of pGEX-4T-1/PaGAI was constructed and transformated into E. coli BL21,and the protein expression was induced by 0.5 mmol · L-1 IPTG and a 91 kD fusion protein was detected by SDS-PAGE. Semi-quantitative RT-PCR analysis showed that PaGAI widely expressed in flower,fruit,leaf and phloem,and PaGAI in the expression of flower and phloem is much stronger than in fruit and leaf.

Key words: Prunus avium, DELLA protein, gene cloning, prokaryotic expression

CLC Number: