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A Chemical-Induced, Seed-Soaking Activation Procedure for Regulated Gene Expression in Rice

机译:化学诱导的浸种激活程序,调控水稻中的基因表达

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Inducible gene expression has emerged as a powerful tool for plant functional genomics. The estrogen receptor-based, chemical-inducible system XVE has been used in many plant species, but the limited systemic movement of inducer β-estradiol in transgenic rice plants has prohibited a wide use of the XVE system in this important food crop. Here, we constructed an improved chemical-regulated, site-specific recombination system by employing the XVE transactivator in combination with a Cre/ loxP -FRT system, and optimized a seed-soaking procedure for XVE induction in rice. By using a gus gene and an hpRNAi cassette targeted for OsPDS as reporters, we demonstrated that soaking transgenic seeds with estradiol solution could induce highly efficient site-specific recombination in germinating embryos, resulting in constitutive and high-level expression of target gene or RNAi cassette in intact rice plants from induced seeds. The strategy reported here thereby provides a useful gene activation approach for effectively regulating gene expression in rice.
机译:诱导型基因表达已成为植物功能基因组学的强大工具。基于雌激素受体的化学诱导系统XVE已在许多植物物种中使用,但是转基因水稻植株中诱导剂β-雌二醇的系统性运动受限,已禁止在这种重要的粮食作物中广泛使用XVE系统。在这里,我们通过将XVE反式激活剂与Cre / loxP -FRT系统结合使用,构建了一种改良的化学调控的位点特异性重组系统,并优化了水稻中XVE诱导的浸种程序。通过使用gus基因和靶向OsPDS的hpRNAi盒作为报告基因,我们证明了用雌二醇溶液浸泡转基因种子可以在发芽胚胎中诱导高效的位点特异性重组,从而导致靶基因或RNAi盒的组成型和高水平表达完整的水稻植株中的诱导种子。因此,本文报道的策略为有效调节水稻中的基因表达提供了有用的基因激活方法。

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