Activation tagging, an efficient tool for functional analysis of the rice genome
Activation tagging, an efficient tool for functional analysis of the rice genome
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DOI：
10.1007/s11103-008-9406-5
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发表时间：
2009-01-01
影响因子：
5.1
通讯作者：
Lu, Tiegang
中科院分区：
文献类型：
--
作者：
Wan, Shuyan;Wu, Jinxia;Lu, Tiegang
在过去的6年里，我们已经产生了约50,000个转基因水稻植株的农杆菌介导的转化方法与pER 38激活标签载体。该载体含有串联排列的双35 S增强子，紧邻T-DNA的右边界。通过逆转录-PCR的表达分析表明，如果基因位于插入的双35 S增强子的7 kb内，则激活效率高。对两代中的部分激活标记和增强子捕获群体（T（0）代中分别为6，000和6，400个株系，T（1）代中分别为36，000和32，000个株系）进行比较田间表型分析，鉴定出约400个显性突变体。具有大叶角的显性突变体（M107）的表征表明，该突变体表型是由CYP 724 B1/D11的增强表达引起的。本文描述的激活标签库是一个有价值的替代工具，水稻基因组的功能分析。
Over the past 6 years, we have generated about 50,000 individual transgenic rice plants by an Agrobacterium-mediated transformation approach with the pER38 activation tagging vector. The vector contains tandemly arranged double 35S enhancers next to the right border of T-DNA. Expression analysis by reverse transcription-PCR indicates that the activation efficiency is high if the genes are located within 7 kb of the inserted double 35S enhancers. Comparative field phenotyping of part of the activation tagging and enhancer trapping populations in two generations (6,000 and 6,400 lines, respectively, in the T(0) generation, and 36,000 and 32,000 lines, respectively, in the T(1) generation) identified about four hundred dominant mutants. Characterization of a dominant mutant with a large leaf angle (M107) suggests that this mutant phenotype is caused by enhanced expression of CYP724B1/D11. The activation tagging pool described in this paper is a valuable alternative tool for functional analysis of the rice genome.