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园艺学报 ›› 2014, Vol. 41 ›› Issue (8): 1689-1698.

• 研究报告 • 上一篇    下一篇

蜜柚泛素蛋白连接酶RING-H2 finger基因的克隆与表达分析

金晓琴,康 振,刘伟娜,潘永娟,王梨嬛,郭卫东,杨 莉*   

  1. (浙江师范大学化学与生命科学学院,浙江金华 321004)
  • 收稿日期:2014-03-04 出版日期:2014-08-25 发布日期:2014-08-25
  • 基金资助:
    国家自然科学基金项目(30900978);浙江省自然科学基金项目(LY14C150001)

Isolation and Expression Analysis of Two Ubiquitin-protein Ligase RING-H2 Finger Genes from Pommelos[Citrus grandis(L.)Osbeck]

JIN Xiao-qin,KANG Zhen,LIU Wei-na,PAN Yong-juan,WANG Li-huan,GUO Wei-dong,and YANG Li*   

  1. (College of Chemistry and Life Science,Zhejiang Normal University,Jinhua,Zhejiang 321004,China)
  • Received:2014-03-04 Online:2014-08-25 Published:2014-08-25

摘要: 在26S泛素蛋白酶体途径介导的蛋白质降解过程中,RING finger蛋白是一类特殊的泛素蛋白连接酶,参与植物多个生长发育及逆境胁迫响应过程。为探明柚RING finger家族成员的序列特征及表达模式,从红肉蜜柚和琯溪蜜柚果实汁胞中分离出两个RING finger基因cDNA片段(CgRHF1CgRHF2),采用实时荧光定量PCR研究其在不同组织及果实发育不同时期的表达模式。结果表明,两个基因均属于RING-H2 finger家族成员,且红肉蜜柚与琯溪蜜柚的两个CgRHF之间碱基序列均无差异;除根组织外,CgRHF1表达水平较CgRHF2高,且在红肉蜜柚茎、叶和花中的表达水平显著高于琯溪蜜柚;CgRHF2在组织及果实发育过程表达水平均较低,但在果实成熟采摘阶段急剧上升。CgRHF1CgRHF2表达模式的差异暗示这两个基因在柚组织及果实发育进程中行使不同的生物学功能,而非功能互补基因。

关键词: 红肉蜜柚, 琯溪蜜柚, RING-H2 finger基因, 克隆, 基因表达

Abstract: RING(Real Interesting New Gene)finger proteins specify the substrates in the 26S ubiquitin-proteasome pathway,and play important roles in plant development and stress responses. However,the sequences,expression characteristics and physiological functions of RING finger genes in pommelo are little known. In this study,two RING finger(CgRHF1 and CgRHF2)cDNA fragments were isolated from Red-fleshed and Guanxi pommelo. The results showed that two CgRHFs were belonged to RING-H2 finger subfamilies,and there is no DNA base difference in the sequences of two cultivars. Quantitative real-time PCR results showed that gene expression of CgRHF1 is significantly higher than CgRHF2,especially in the stems,leaves and flowers of red-fleshed pommelo. The expression levels of CgRHF2 were relatively low in the tissues and fruit development of two pommelo cultivars,but dramatically increased at the fruit mature stage. This study has implied that CgRHF1 and CgRHF2 may play different roles in the tissues and fruit development of Red-fleshed and Guanxi pommelo,and can not functionally complement.

Key words: red-fleshed pommelo, Guanxi pommelo, RING-H2 finger gene, clone, gene expression

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