水稻泛素连接酶基因OsRING6的克隆及表达分析  

Cloning and Expression Analysis of the Rice Ubiquitin Ligase Gene OsRING6

作　　者：范锡麟[1] 杜强 杨珍珍 何艳冰 王国梁[1,2] 王志龙 

机构地区：[1]湖南省作物种质创新与资源利用重点实验室/湖南农业大学农学院,长沙410128 [2]Department of Plant Pathology, The Ohio State University, Columbus 43210

出　　处：《植物遗传资源学报》2016年第2期337-343,共7页Journal of Plant Genetic Resources

基　　金：国家自然科学基金项目(31571995);湖南省高校科技创新团队项目(2012);湖南省研究生科研创新项目(CX2014B288)

摘　　要：RING(Really Interesting New Gene)型泛素连接酶在水稻生长、发育、生物和非生物胁迫中发挥着重要作用。根据水稻基因组信息,利用RT-PCR技术从粳稻日本晴中克隆了OsRING6基因的CDS序列。该基因的全长c DNA为1355 bp,包括长度为918 bp的CDS,编码一个含305个氨基酸的蛋白,预测该蛋白的分子量和等电点分别为32.96 k D和3.49,与籼稻(CT832014)和短花药野生稻(XM_006659599)中同源蛋白的序列相似性最高,分别为97%和83%。IPTG诱导表达的MBPOsRING6重组蛋白的表观分子量约为100 k D,比理论值76 k D大。启动子元件分析表明,其含有与干旱胁迫、光反应、ABA和GA信号传导相关的调控元件。Real-time PCR结果表明,该基因在水稻根、茎、叶、叶鞘、花和种子中都有表达,在萌发后4 d的种子中表达量最高,花中的表达量最低,且该基因的表达受到盐和ABA的诱导及PEG的抑制。OsRING6基因的克隆为水稻抗逆分子育种和功能分析提供了优质基因资源。RING（ Really Interesting New Gene） ubiquitin ligases play significant roles in rice,such as growth,development,resistance to biotic and abiotic stresses. In this study,we cloned the RING finger gene（ OsRING6） from the Japonica cultivar Nipponbare（ Oryza sativa） based on the sequence analysis of the rice genome and the RT-PCR method. The OsRING6 full c DNA length was 1355 bp and contained a CDS of 918 bp nucleotides,and encoded a protein of 305 amino acids with the deduced molecular weight of 32. 96 k D and p I value of 3. 49. The similarity comparison revealed that the CDS sequence of OsRING6 shared 97 % and 83 %homology with Oryza sativa Indica（ CT832014） and Oryza brachyantha（ XM_006659599）,respectively. The relative molecular weight of the MBP-OsRING6 recombinant protein induced by IPTG was about 100 k D,which was much bigger than deduced value of 76 k D. Promoter analysis indicated that it contained several regulatory elements related to drought stress,light response,ABA and GA signal transduction. Real-time PCR experiments showed that OsRING6 was expressed mainly in root,stem,leaves,leaf sheath,flowers and seed,and its expression level was the highest in germinated seeds and the lowest in flowers among these detected tissues.We also found that OsRING6 could be induced when treated with salt and ABA and inhibited by PEG. These data suggestted that OsRING6 might be involved in abiotic stresses such as slat and drought,and could be usedfor further functional study and breeding in rice.

关 键 词：水稻 泛素连接酶 RING 启动子 表达分析 

分 类 号：S511.22[农业科学—作物学]