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Ac/Ds转座系统在水稻转化群体中的转座活性及转座子插入位点旁侧序列的分析
引用本文:朱正歌,付亚萍,肖晗,胡国成,斯华敏,于永红,孙宗修.Ac/Ds转座系统在水稻转化群体中的转座活性及转座子插入位点旁侧序列的分析[J].Acta Botanica Sinica,2003,45(1):102-107.
作者姓名:朱正歌  付亚萍  肖晗  胡国成  斯华敏  于永红  孙宗修
作者单位:中国水稻研究所农业部水稻生物学重点实验室,中国水稻研究所农业部水稻生物学重点实验室,中国水稻研究所农业部水稻生物学重点实验室,中国水稻研究所农业部水稻生物学重点实验室,中国水稻研究所农业部水稻生物学重点实验室,中国水稻研究所农业部水稻生物学重点实验室,中国水稻研究所农业部水稻生物学重点实验室 杭州 310006,河北师范大学生命科学学院.石家庄 050016,杭州 310006,杭州 310006,杭州 310006,杭州 310006,杭州 310006,杭州 310006
基金项目:国家自然科学基金重点项目(39789020),国家重大基础研究发展规划项目(G199901160)~~
摘    要:利用本实验室构建的转Ac(Ac TPase)及Ds(Dissociation)的水稻(Oryza sativa L.)转化群体,配置了Ae×Ds的杂交组合354个。检测了转基因植株的T-DNA插入位点右侧旁邻序列,研究了Ac/Ds转座系统在水稻转化群体中的转座活性。结果表明,有些转化植株T-DNA插入位点相同或相距很近,插入位点互不相同的占65.4%。检测到T-DNA可插入到编码蛋白的基因中。在Ac×Ds的F2代中,Ds因子的转座频率为22.7%。对Ac×Ds杂交子代中Ds因子旁侧序列的分析,进一步表明了Ds因子在水稻基因组中的转座活性,除了从原插入位点解离并转座到新的位点之外,还有复制——转座和小完全切离等现象。获得的旁侧序列中,有些序列与GenBank中的数据没有同源性,目前有2个DNA片段在GenBank登录。探讨了构建转座子水稻突变体库进行水稻功能基因组学研究的策略。

关 键 词:Ac/Ds转座系统  水稻  转化群体  转座活性  转座子  插入位点  旁侧序列

Ac/Ds Transposition Activity in Transgenic Rice Population and DNA Flanking Sequence of Ds Insertion Sites
Authors:ZHU Zheng-Ge  FU Ya-Ping  XIAO Han  HU Guo-Cheng  SI Hua-Min  YU Yong-Hong  SUN Zong-Xiu
Abstract:The Agrobacterium mediated transgenic rice ( Oryza saliva L.) population with inserts of maize transposon Activator/Dissociation (Ac/Ds) was investigated. DNA sequences flanking the T-DNA were analyzed with inverse PCR. Results showed that 65.4% of the T-DNA was integrated in different locations of rice genome, and some T-DNA flanking sequences were located on certain chromosomes. A number of T-DNA was found to have inserted into protein coding regions. In order to induce transposition of the inserted Ds elements, 354 crosses of Ac x Ds and Ds x Ac were constructed. The excision frequency of Ds element trans-activated by Ac transposase was 22.7% in the F2 populations, and the transposition was confirmed with analyses of DNA sequences flanking the Ds elements. In addition to the transposition due to "cut-paste" mechanism, Ds can replicate itself and integrate into a new locus, and inaccurate excisions were also found. A proportion of DNA segments flanking the Ds elements showed no homologies to sequences published in GenBank, of which two were registered under the accession numbers AF355153 and AF355770. The strategy of using transposon tagging for rice genomics study was discussed.
Keywords:transposon Ac/Ds  transposition activity  flanking sequence  Oryza saliva  
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