Transposon display for active DNA transposons in rice
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概要
- 論文の詳細を見る
Transposon display (TD) is a powerful technique to identify the integration site of transposons in gene tagging as a functional genomic tool for elucidating gene function. Although active endogenous DNA transposons have been used extensively for gene tagging in maize, only two active endogenous DNA transposons in rice have been identified, the 0.43-kb element mPing of the MITE family and the 0.6-kb nDart element of the hAT family. The nDart transposition was shown to be induced by crossing with a line containing its autonomous element aDart and stabilized by segregating aDart under natural growth conditions, while mPing-related elements were shown to transpose in cultured cells, plants regenerated from an anther culture, and γ-ray-irradiated plants. No somaclonal variation should occur in nDart-promoted gene tagging because no tissue culture was involved in nDart activation. As an initial step to develop an effective tagging system using nDart in rice, we tried to visualize GC-rich nDart-related elements comprising 18 nDart-related sequences of 0.6-kb and 63 nDart-related elements longer than 2 kb in Nipponbare by TD. Comparing the observed bands in TD with the anticipated virtual bands of the nDart-related elements based upon the available rice genome sequence, we have improved our TD protocol by optimizing the PCR amplification conditions and are able to visualize approximately 87% of the anticipated bands produced from the nDart-related elements. To compare the visualization efficiency of these nDart-related elements with that of 50 mPing elements and a unique Ping sequence in Nipponbare, we also tried to visualize the mPing-related elements; all mPing-related elements are easily visualized. Based on these results, we discuss the parameters affecting the visualization efficiencies of these rice DNA transposons. We also discuss the utilization of nDart elements in gene tagging for functional genomics in rice.
- 日本遺伝学会の論文
- 2007-04-25
著者
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Ishikawa Naoko
National Inst. Natural Sci. Okazaki Jpn
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Ishikawa Naoko
National Institute For Basic Biology
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Tsugane Kazuo
National Inst. For Basic Biology
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Tsugane Kazuo
National Institute For Basic Biology
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IIDA Shigeru
National Institute for Basic Biology
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Maekawa Masahiko
Institute of Plant Science and Resources, Okayama University
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Maekawa Masahiko
Research Institute for Science and Technology, Kinki University
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TAKAGI Kyoko
Graduate School of Agriculture, Hokkaido University
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Takagi Kyoko
Graduate School Of Agriculture Hokkaido University
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Iida Shigeru
National Inst. For Basic Biology
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Maekawa Masahiko
Research Institute For Bioresources Okayama University
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Iida Shigeru
Graduate School Of Pharmaceutical Sciences Univ. Of Shizuoka
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