Plant synthetic biology requires precise characterization of genetic elements to construct complex genetic circuits that can improve plant traits or confer them with new characteristics. Transcriptional reporter assays are essential to quantify the effect of gene expression regulator elements. Additionally, transcriptional reporter systems are a key tool in understanding control of gene expression in biology. In this work, we construct and characterize a dual color luciferase ratiometric reporter system that possesses several advantages over currently used reporters. It is ratiometric, thus reducing variability and increasing consistency between experiments; it is fast, as both reporters can be measured at the same time in a single reaction, and it is less expensive to perform than current dual luciferase reporter assays. We have validated our system quantifying the transcriptional capability of a panel of promoters and terminators commonly used in synthetic biology with a broad range of expression magnitudes, and in a biologically relevant system, nitrate response.

中文翻译：

---

用于植物转录调控元件快速表征的比率双色荧光素酶报告基因

植物合成生物学需要对遗传元件进行精确表征，以构建复杂的遗传电路，从而改善植物性状或赋予它们新的特性。转录报告分析对于量化基因表达调控元件的作用至关重要。此外，转录报告系统是理解生物学中基因表达控制的关键工具。在这项工作中，我们构建并表征了双色荧光素酶比率报告系统，该系统比目前使用的报告系统具有多种优势。它是比例式的，因此减少了变异性并增加了实验之间的一致性；它速度很快，因为可以在一个反应​​中同时测量两个报告基因，并且比当前的双荧光素酶报告基因检测更便宜。我们已经验证了我们的系统，量化了合成生物学中常用的一组启动子和终止子的转录能力，具有广泛的表达量，以及在生物学相关系统中的硝酸盐反应。