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首页> 外文期刊>Genes & Genetic Systems >Transposon display for active DNA transposons in rice.
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Transposon display for active DNA transposons in rice.

机译：转座子展示水稻中活性DNA转座子。

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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 gamma -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..
机译：转座子展示（TD）是一种强大的技术，可在基因标记中鉴定转座子的整合位点，作为阐明基因功能的功能基因组工具。尽管活性内源DNA转座子已广泛用于玉米中的基因标记，但仅在水稻中鉴定出两个活性内源DNA转座子，即MITE家族的0.43-kb元件mPing和hAT家族的0.6-kb nDart元件。显示nDart转座是通过与包含其自主元件aDart的品系杂交而诱导的，并通过在自然生长条件下分离aDart来稳定，而显示mPing相关元件可在培养细胞，从花药培养物再生的植物和γ中转座。射线照射的植物。在nDart促进的基因标记中不应发生体细胞克隆变异，因为nDart激活中不涉及组织培养。作为开发在水稻中使用nDart的有效标签系统的第一步，我们尝试通过TD可视化在日本晴中可视化包含18个0.6kb的nDart相关序列和63个长于2 kb的nDart相关元素的富含GC的nDart相关元素。 。根据可用的水稻基因组序列，将TD中观察到的条带与nDart相关元件的预期虚拟条带进行比较，我们通过优化PCR扩增条件改进了TD方案，并能够可视化产生约87％的预期条带来自与nDart相关的元素。为了将这些与nDart相关的元素的可视化效率与50个mPing元素的可视化效率以及Nipponbare中唯一的Ping序列进行比较，我们还试图对与mPing相关的元素进行可视化。所有与mPing相关的元素均易于查看。基于这些结果，我们讨论了影响这些水稻DNA转座子可视化效率的参数。我们还讨论了nDart元素在水稻功能基因组学的基因标签中的应用。

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