Scientia Agricultura Sinica ›› 2012, Vol. 45 ›› Issue (7): 1347-1354.doi: 10.3864/j.issn.0578-1752.2012.07.012

• HORTICULTURE • Previous Articles     Next Articles

Cloning and Prokaryotic Expression of MdRGL Gene from Spur-Type Apple (Malus domestica Borkh.)

 SONG  Yang, ZHANG  Yan-Min, LIU  Mei-Yan, WANG  Chuan-Zeng, LIU  Jin, FENG  Shou-Qian, WANG  Yan-Ling, CHEN  Xue-Sen   

  1. 山东农业大学园艺科学与工程学院/作物生物学国家重点实验室，山东泰安 271018
  • Received:2011-11-01 Online:2012-04-01 Published:2012-02-20

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Abstract: 【Objective】Cloning, sequence analysis of the DELLA genes from Malus domestica Borkh and its expression it in E. coli were conducted to further explore the relationship between DELLA and mutation shoot of spur type bud sport apple.【Method】The MdRGL cDNA fragments amplified from Malus domestica Borkh by reverse transcription PCR (RT-PCR) and then the cloned genes of MdRGL were inserted into vector pGEX-4T-1. The recombinant plasmids pGEX-4T-MdRGL were expressed in a prokaryotic expression system after its transformation into E. coli BL21 (DE3).【Result】 Five DELLA genes, MdRGL1a/b, MdRGL2a/b and MdRGL3b were isolated. Molecular structure analysis revealed that these five genes exhibited the typical structures of the DELLA and VHYNP domain except MdRGL3b. The results of SDS-PAGE demonstrated that the expressed proteins were consistent with the size of expected protein in the prokaryotic expression system.【Conclusion】The five DELLA genes of spur-type apple were cloned and successfully expressed in E. coli. This study will provide a foundation for studying the function of the target protein.

Key words: spur-type apple, DELLA protein, gene clone, sequence analysis, prokaryotic expression

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