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

作　　者：Zhimin He Bin Liu Xu Wang Mingdi Bian Reqing He Jindong Yan Ming Zhong Xiaoying Zhao Xuanming Liu 

机构地区：[1]Hunan Province Key Laboratory of Plant Functional Genomics and Developmental Regulation, College of Biology, Hunan University, Changsha 410082, China [2]Department of Molecular, Cell & Developmental Biology, University of California, Los Angeles, CA 90095, USA [3]Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing 100081, China [4]College of Plant Sciences, Jilin University, Changchun 130062, China [5]State Key Laboratory of Chemo/Biosensing and Chemometrics, Hunan University, Changsha 410082, China

出　　处：《Journal of Genetics and Genomics》2016年第4期207-215,共9页遗传学报（英文版）

基　　金：supported by the National Natural Science Foundation of China(Grant No.31171176);the Cooperative Innovation Center of Engineering and New Products for Developmental Biology of Hunan Province(Grant No.20134486)

摘　　要：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.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.

关 键 词：CO-EXPRESSION Dual-transgene vector pDT1 pDT7 pDT7G 

分 类 号：Q943.2[生物学—植物学]