Specific expression of DR5 promoter in rice roots using a tCUP derived promoter-reporter system.
Specific expression of DR5 promoter in rice roots using a tCUP derived promoter-reporter system.
复制标题
使用 tCUP 衍生的启动子-报告基因系统进行 DR5 启动子在水稻根中的特异性表达
DOI：
10.1371/journal.pone.0087008
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Chen J
中科院分区：
文献类型：
--
作者：
Zhou J;Yu F;Wang X;Yang Y;Yu C;Liu H;Cheng Y;Yan C;Chen J
由于插入位点的位置效应或用于表达可选择标记基因的启动子/增强子元件引起的转基因表达的变化具有复杂的表型特征和引起的误解。我们建立了水稻报告基因系统，分析了载体结构、间隔区和选择标记基因启动子对无启动子GUS报告基因和DR5启动子表达的影响。我们的结果表明，在反向35S启动子和GUS报告基因之间插入间隔区可以减少报告基因的泄漏表达，但不能阻止DR5：：GUS的非特异性表达。即使当35S启动子用于表达可选标记时，将可选标记单元与GUS报告基因从头到尾堆叠有助于DR5启动子下的GUS报告基因的特异性表达。与35S相比，通过使用TCUP衍生的启动子(TCUP1)进行选择，DR5：：GUS在根尖分生组织的根冠、静止中心和木质部细胞中得到了快速而独特的表达，这与拟南芥中敏感的DR5突变体(DR5rev)获得的模式相似。这些数据表明，TCUP启动子在防止水稻中的增强子-启动子相互作用方面具有与拟南芥相同的保守性质，也表明在水稻的根中存在类似的远端生长素最大值。因此，基于TCUP启动子的选择系统为转基因在水稻中的特异性表达提供了新的策略。
Variation of transgene expression caused by either position effect at the insertion site or the promoter/enhancer elements employed for the expression of selectable marker genes has complicated phenotype characterization and caused misinterpretation. We have developed a reporter system in rice to analyze the influence of vector configuration, spacer and selectable marker gene promoter on the expression of the promoterless GUS reporter and DR5 promoter. Our results indicate that a spacer inserted between the reversed 35S promoter and the GUS reporter could reduce leaky expression of the reporter but was unable to block the nonspecific expression of DR5::GUS. Stacking the selectable marker unit in head to tail with the GUS reporter aided the gene specific expression of the GUS reporter under the DR5 promoter even when the 35S promoter is used for expression of the selectable marker. Compared to 35S under this configuration, a quick and distinctive expression of DR5::GUS was observed in the root cap, quiescent center and xylem cells in the root apical meristem by using the tCUP derived promoter (tCUP1) for selection, that is similar to the pattern obtained by a sensitive DR5 variant (DR5rev) in Arabidopsis. These data suggest a conserved property of the tCUP promoter in preventing enhancer-promoter interactions in rice as it does in Arabidopsis, and also demonstrate that an analogous distal auxin maximum exists in roots of rice. Therefore, the tCUP promoter based selection system provides a new strategy for specific expression of transgenes in rice.
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通讯作者：
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作者：
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通讯作者：
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作者：
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