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    Please use this identifier to cite or link to this item: http://asiair.asia.edu.tw/ir/handle/310904400/7959


    Title: A rice gene activation/knockout mutant resource for high throughput functional genomics.
    Authors: Yue-Ie Hsing;Chyr-Guan Chern;Ming-Jen Fan;Po-Chang Lu;Ku-Ting Chen;Shuen-Fang Lo;Peng-Kai Sun;Shin-Lon Ho;Kuo-Wei Lee;Yi-Chieh Wang;Wen-Li Huang;Swee-Suak Ko;Shu Chen;Jyh-Long Chen;Chun-I Chung;Yao-Cheng Lin;Ai-Ling Hour;Yet-Walt Wang;Ya-Chi Chang;Min-Wei Tsai;Yi-Show Lin;Yin-Chin Chen;Hsing-Mu Yen;Charng-Pei Li;Chiu-Kai Wey;Ching-Shan Tseng;Ming-Hsing Lai;Sheng-Chung Huang;Liang-Jwu Chen;Su-May Yu
    Contributors: Department of Biotechnology
    Date: 2007
    Issue Date: 2010-03-15 16:10:44 (UTC+8)
    Publisher: Asia University
    Abstract: Using transfer DNA (T-DNA) with functions of gene trap and gene knockout and activation tagging, a mutant population containing 55,000 lines was generated. Approximately 81% of this population carries 1-2 T-DNA copies per line, and the retrotransposon Tos17 was mostly inactive in this population during tissue culture. A total of 11,992 flanking sequence tags (FSTs) have been obtained and assigned to the rice genome. T-DNA was preferentially ( approximately 80%) integrated into genic regions. A total of 19,000 FSTs pooled from this and another T-DNA tagged population were analyzed and compared with 18,000 FSTs from a Tos17 tagged population. There was difference in preference for integrations into genic, coding, and flanking regions, as well as repetitive sequences and centromeric regions, between T-DNA and Tos17; however, T-DNA integration was more evenly distributed in the rice genome than Tos17. Our T-DNA contains an enhancer octamer next to the left border, expression of genes within genetics distances of 12.5 kb was enhanced. For example, the normal height of a severe dwarf mutant, with its gibberellin 2-oxidase (GA2ox) gene being activated by T-DNA, was restored upon GA treatment, indicating GA2ox was one of the key enzymes regulating the endogenous level of GA. Our T-DNA also contains a promoterless GUS gene next to the right border. GUS activity screening facilitated identification of genes responsive to various stresses and those regulated temporally and spatially in large scale with high frequency. Our mutant population offers a highly valuable resource for high throughput rice functional analyses using both forward and reverse genetic approaches.
    Relation: Plant Molecular Biology 63(3):351-64
    Appears in Collections:[生物科技學系] 期刊論文

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