Pasin Fabio, Bedoya Leonor C, Bernabé-Orts Joan Miquel, Gallo Araíz, Simón-Mateo Carmen, Orzaez Diego, García Juan Antonio
Centro Nacional de Biotecnología (CNB-CSIC) , Darwin 3, 28049 Madrid, Spain.
Instituto de Biología Molecular y Celular de Plantas (IBMCP, CSIC-UPV) , Camino de Vera s/n, 46022 Valencia, Spain.
ACS Synth Biol. 2017 Oct 20;6(10):1962-1968. doi: 10.1021/acssynbio.6b00354. Epub 2017 Jun 28.
Improved plants are necessary to meet human needs. Agrobacterium-mediated transformation is the most common method used to rewire plant capabilities. For plant gene delivery, DNA constructs are assembled into binary T-DNA vectors that rely on broad host range origins for bacterial replication. Here we present pLX vectors, a set of mini binary T-DNA plasmids suitable for Type IIS restriction endonuclease- and overlap-based assembly methods. pLX vectors include replicons from compatible broad host range plasmids. Simultaneous usage of pBBR1- and RK2-based pLX vectors in a two-plasmid/one-Agrobacterium strain strategy allowed multigene delivery to plants. Adoption of pLX vectors will facilitate routine plant transformations and targeted mutagenesis, as well as complex part and circuit characterization.
改良植物对于满足人类需求至关重要。农杆菌介导的转化是用于改变植物能力的最常用方法。对于植物基因传递，DNA构建体被组装成二元T-DNA载体，这些载体依赖于广泛宿主范围的复制起点进行细菌复制。在此，我们展示了pLX载体，这是一组适用于IIS型限制性内切酶和重叠组装方法的微型二元T-DNA质粒。pLX载体包含来自兼容的广泛宿主范围质粒的复制子。在双质粒/单农杆菌菌株策略中同时使用基于pBBR1和RK2的pLX载体可实现向植物的多基因传递。采用pLX载体将有助于常规的植物转化和靶向诱变，以及复杂部件和电路的表征。
