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

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• 作者：Zhimin He^a ; ^b ; Bin Liu^c ; Xu Wang^b ; Mingdi Bian^d ; Reqing He^a ; Jindong Yan^a ; Ming Zhong^a ; Xiaoying Zhao^a ; ^{xiaoyzhao@hnu.edu.cn" class="auth_mail" title="E-mail the corresponding author} ; Xuanming Liu^a ; ^e ; ^{xmL05@hnu.edu.cn" class="auth_mail" title="E-mail the corresponding author}
• 关键词：Co-expression ; Dual-transgene vector ; pDT1 ; pDT7 ; pDT7G
• 刊名：Journal of Genetics and Genomics
• 出版年：2016
• 出版时间：20 April 2016
• 年：2016
• 卷：43
• 期：4
• 页码：199-207
• 全文大小：1836 K

文摘

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 Arabidopsis thaliana. The co-expression efficiency of two genes from the pDT1 and pDT7G 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.