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园艺学报 ›› 2006, Vol. 33 ›› Issue (2): 239-243.

• 研究论文 • 上一篇    下一篇

苹果apetala2同源基因的克隆和转化研究

周盛梅1;宿红艳2;王 磊2;张宪省3;束怀瑞1*
  

  1. (1 山东农业大学园艺科学与工程学院, 泰安271018; 2 烟台师范学院生命科学学院, 烟台264025; 3 山东农业大学生命科学学院, 泰安271018)
  • 收稿日期:2005-03-04 修回日期:2006-02-02 出版日期:2006-04-25 发布日期:2006-04-25

Cloning and Transformation of apetala2 Homologues Genes from Apple

Zhou Shengmei1;Su Hongyan2;Wang Lei2;Zhang Xiansheng3;and Shu Huairui1*

  

  1. (1 Institute of Horticulture Science and Engineering, Shandong Agricultural University, Tai'an 271018, China; 2College of L ife Sciences, Yantai N orm al University, Yantai 264025, China; 3College of L ife Sciences, Shandong Agricultural University, Tai'an 271018, China)
  • Received:2005-03-04 Revised:2006-02-02 Online:2006-04-25 Published:2006-04-25

摘要: 利用同源克隆的方法从苹果的花芽中分离出apetala2的同源基因MAP2MAP2全长2 212 bp,编码549个氨基酸。分析表明, MAP2具有AP2家族典型的结构域, 是苹果的AP2同源基因。Southern杂交结果表明, MAP2在基因组中以低拷贝形式存在。采用RT-PCR的方法分析MAP2在不同组织中的表达,结果显示, MAP2在苹果营养组织、花芽以及不同花器官中均有表达, 与拟南芥的AP2、矮牵牛的PhAP2A的表达模式一致。为确定MAP2在苹果中的生物学功能, 构建了35S∶MAP2正义表达载体, 并对‘皇家嘎啦’苹果进行了农杆菌介导的遗传转化。

关键词: 苹果, apetala2, 基因克隆, 表达分析, 转基因

Abstract: MAP2, with homology to Arabidopsis AP2, was isolated from the flower buds of apple (Malus domestica Borkh). MAP2 cDNA is 2 212 bp long and encodes a 549 amino acid protein. Like other AP2 genes, MAP2 encodes a protein containing two distinct AP2 domain, a transcription-activating domain and aputative nuclear location signal. This indicates thatMAP2 is a homologue of AP2 in app le. Although the homolog among the three proteins in the putative nuclear location signal is 100% , MAP2 shares only 45.1% and 55.2% similarity with AP2 and PhAP2A , respectively. This reflects the diversity in the function of AP2 gene family excep t for acting in the nuclear. RT-PCR analysis showed that MAP2 is expressed in floral buds, allfloral organs and vegetative organs, which is similar to the exp ression pattern of AP2 and PhAP2A. To study the function of MAP2 in apple, we introduced the sense MAP2 under the control of 35S promoter of Cauliflower mosaic virus into Royal Gala plants.

Key words: Apple, apetala2, Gene cloning, Expression, Transgenic