Table 1_pTARGEX vectors: a versatile toolbox for plant-based protein expression.docx

The ability to express heterologous proteins in specific plant tissues and subcellular compartments is critical for advancing plant-based biotechnologies and biopharming research. Here, we present the “pTARGEX” series, a versatile toolkit designed to enable targeted protein expression within the leaves and roots of plants. These vectors incorporate subcellular targeting sequences to direct proteins of interest to the apoplast (pTARGEX-APO), cytoplasm (pTARGEX-CYT), chloroplast (pTARGEX-CHL), endoplasmic reticulum (pTARGEX-ER), or vacuole (pTARGEX-VAC). Streamlining workflow, the pTARGEX vectors encode an mCherry dropout cassette with a SapI restriction enzyme site to enable red/white screening in Escherichia coli, and an eGFP reporter gene that is constitutively expressed in planta to serve as an inbuilt marker for monitoring of successful transformation. This innovative vector series offers users an accessible, versatile platform for plant-based heterologous protein production, facilitating research and applications in systems biology, synthetic biology, and plant biotechnology.

在特定植物组织与亚细胞区室中表达异源蛋白的能力，对于推动植物生物技术及植物生物制药研究的发展具有关键意义。在此我们推出pTARGEX系列载体工具包，该工具包可实现目标蛋白在植物叶片与根系中的靶向表达。该系列载体搭载亚细胞靶向序列，可将目标蛋白分别引导至质外体（apoplast）、细胞质（cytoplasm）、叶绿体（chloroplast）、内质网（endoplasmic reticulum）或液泡（vacuole）中。为简化实验流程，pTARGEX载体携带带有SapI限制性酶切位点的mCherry筛选剔除盒，可在大肠杆菌（Escherichia coli）中实现红/白菌落筛选；同时还搭载了组成型表达的eGFP（enhanced green fluorescent protein）报告基因，可作为植物体内遗传转化成功与否的内置检测标记。该创新载体系列为用户提供了便捷通用的植物源异源蛋白表达平台，可助力系统生物学、合成生物学及植物生物技术领域的研究与应用。