华南理工大学学报(自然科学版) ›› 2011, Vol. 39 ›› Issue (5): 143-148.doi: 10.3969/j.issn.1000-565X.2011.05.025

• 生物工程 • 上一篇    下一篇

盐芥TOR 基因的克隆及对盐芥生长发育的影响

王臻 吴春霞 扈玉婷 张慧 赵彦修   

  1. 山东师范大学 山东省逆境植物重点实验室,山东 济南 250014
  • 收稿日期:2010-11-18 出版日期:2011-05-25 发布日期:2011-04-01
  • 通信作者: 赵彦修(1954-),男,教授,博士生导师,主要从事植物耐逆研究.E-mail:arabidopsis@sdnu.edu.cn E-mail:musicpianowz@163.com
  • 作者简介:王臻(1983-) ,女,博士生,主要从事植物耐逆分子生物学研究.
  • 基金资助:

    国家“973”计划项目( 2006CB100104) ; 国家“863”计划项目( 2007AA021402)

Cloning of TOR from Thellungiella halophila and Its Role in Plant Growth

Wang Zhen  Wu Chun-xia  Hu Yu-ting  Zhang Hui  Zhao Yan-xiu   

  1. Shandong Provincial Key Laboratory of Plant Stress Research,Shandong Normal University,Jinan 250014,Shandong,China
  • Received:2010-11-18 Online:2011-05-25 Published:2011-04-01
  • Contact: 赵彦修(1954-),男,教授,博士生导师,主要从事植物耐逆研究.E-mail:arabidopsis@sdnu.edu.cn E-mail:musicpianowz@163.com
  • About author:王臻(1983-) ,女,博士生,主要从事植物耐逆分子生物学研究.
  • Supported by:

    国家“973”计划项目( 2006CB100104) ; 国家“863”计划项目( 2007AA021402)

摘要: 雷帕霉素靶蛋白( TOR) 在调节动植物的生长及新陈代谢方面起重要作用.为进一步研究植物中TOR 的作用,对盐芥TOR 基因进行克隆,采用聚合酶链式反应和cDNA末端快速扩增技术等分子生物学方法获得盐芥TOR 基因的cDNA 全长及基因组全长,构建了ThTOR 基因沉默植物表达载体并成功获得盐芥ThTOR 基因沉默的转基因株系.结果表明: ThTOR cDNA 全长为7 836 bp,编码2 479 个氨基酸,蛋白序列与拟南芥同源性高达97. 5%,基因组全长19162 bp; 盐芥ThTOR 基因沉默的转基因株系具有生长缓慢、花及种子的发育受阻的明显表型.

关键词: TOR, 盐芥, 克隆, 遗传转化, RNA 干扰

Abstract:

Mammalian target of rapamycin ( TOR) plays an important role in the growth regulation and metabolism of animals and plants. In order to reveal the function of TOR in plants,TOR gene of Thellungiella halophila ( T. halophila) was cloned,and the full lengths of ThTOR cDNA and genomic DNA were obtained by polymerase chain reaction ( PCR) and rapid amplification of cDNA ends ( RACE) . Then,RNAi vector ThTOR-RNAi was constructed
and the transgenic lines were obtained by transforming ThTOR-RNAi into T, halophila. The results show that the full-length ThTOR cDNA contains 7836 base pairs and encodes a protein with 2479 amino acids,that a high degree of homology ( 97. 5%) exists between the proteins encoded by ThTOR and AtTOR,that the full-length ThTOR genomic DNA is of 19 162 base pairs,and that obvious phenotypes of slow growth as well as flowers and seeds abortion occur in T. halophila ThTOR-RNAi transgenic lines.

Key words: mammalian target of rapamycin, Thellungiella halophila, cloning, genetic transformation, RNA interference