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林业科学 ›› 2010, Vol. 46 ›› Issue (2): 45-50.doi: 10.11707/j.1001-7488.20100208

• 论文 • 上一篇    下一篇

绿竹BoSUT2基因的分子特征与亚细胞定位

高志民1 杨学文1 彭镇华1,2 李雪平1 牟少华1 马艳军1   

  1. 1.国际竹藤网络中心 国家林业局竹藤科学与技术重点开放实验室 北京 100102; 2.中国林业科学研究院林业研究所 北京 100091
  • 收稿日期:2009-07-06 修回日期:1900-01-01 出版日期:2010-02-25 发布日期:2010-02-25
  • 通讯作者: 彭镇华

Molecular Characterization and Subcellular Localization of BoSUT2 from Bambusa oldhamii

Gao Zhimin1,Yang Xuewen1,Peng Zhenhua1,2,Li Xueping1,Mu Shaohua1,Ma Yanjun1   

  1. 1.Key Laboratory of Bamboo and Rattan Science and Technology of State Forestry Administration International Center for Bamboo and Rattan Beijing 100102; 2.Research Institute of Forestry, Chinese Academy of Forestry Beijing 100091
  • Received:2009-07-06 Revised:1900-01-01 Online:2010-02-25 Published:2010-02-25

摘要:

植物蔗糖转运蛋白( sucrose transporter, SUT) 作为一种功能性蛋白, 在蔗糖运输以及蔗糖特异信号感应过程中发挥着重要的生理功能。采用RT-PCR方法从绿竹中分离1个新的编码蔗糖转运蛋白基因,cDNA长1 668 bp,编码555个氨基酸,命名为BoSUT2(注册号: EU247931)。序列分析表明,BoSUT2与其他单子叶植物的蔗糖转运蛋白有着较高的一致性,其中与绿竹BoSUT1一致性最高,达92.3%。编码蛋白二级结构预测分析显示,BoSUT2具有糖运转子保守域和跨膜结构域,属于膜蛋白。组织特异性表达检测表明,BoSUT2在叶片和叶鞘中的表达丰度较高且比较接近,在根中有微弱表达。将BoSUT2置于35S启动子之下,构建了含GFP的融合表达载体,并转化烟草悬浮细胞,观察结果表明,BoSUT2主要定位到细胞质膜上。

关键词: 绿竹, 蔗糖转运蛋白基因, 组织特异性表达, 亚细胞定位

Abstract:

Sucrose transporter is a functional protein which plays an important role in transporting sucrose and sensing specific sucrose signals. A sucrose transporter gene, named as BoSUT2 (GenBank accession number: EU247931), has been isolated from the first strand of Bambusa oldhamii cDNA through RT-PCR method. The cDNA is 1 668 bp long and contains an open reading frame which encodes a polypeptide of 555 amino acids. The result of a homology analysis shows that the deduced BoSUT2 is highly homologous to other SUT proteins from various monocotyledonous species. The similarity with BoSUT1 from B. oldhamii is the highest, reaching to 92.3%. Secondary structure prediction analysis shows that BoSUT2 has trans-membrane domains and sucrose transporter conserved domains, which indicates that BoSUT2 belongs to membrane proteins. BoSUT2 is expressed highly in leaf and sheath, and weakly in root as shown in tissue specific expression tests. BoSUT2 gene is constructed in the GFP fusion expression vector under the control of CaMV 35S promoter; and then transferred into tobacco suspension cells. The results of subcellular localization analysis further demonstrate that BoSUT2 proteins are mainly positioned on the cytoplasmic membrane.

Key words: Bambusa oldhamii, sucrose transporter gene, tissue specific expression, subcellular localization