Construction and Validation of a Dual-Transgene Vector System for Stable Transformation in Plants

In this study,we constructed dual-transgene vectors(pDT1,pDT7,and pDT7G) that simultaneously co-expressed two genes in plants.ACTIN2 and UBQ10 promoters were used to control the expression of these two genes.The 4×Myc.3×HA,and 3×Flag reporter genes allowed for the convenient identification of a tunable co-expression system in plants,whereas the dexamethasone(Dex) inducible reporter gene C-terminus of the glucocorticoid receptor(cGR) provided Dex-dependent translocation of the fusion gene between the nucleus and cytoplasm.The function of pDT vectors was validated using four pairwise genes in Nicotiana benthamiana or Anihidopsis thaliana.The co-expression efficiency of two genes from the pDT1 and pDT7 G vectors was 35%and 42%,respectively,which ensured the generation of sufficient transgenic materials.These pDT vectors are simple,reliable,efficient,and time-saving tools for the co-expression of two genes through a single transformation event and can be used in the study of protein-protein interactions or multi-component complexes.