Inducible expression in plants by virus-mediated transgene activation

• Published in: Transgenic research Vol. 14; no. 4; pp. 407 - 416
• Main Authors: Hull, A.K, Yusibov, V, Mett, V
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer 01.08.2005; Springer Nature B.V
• Subjects: agroinfiltration; beta-glucuronidase; Biological and medical sciences; Biotechnology; Blotting, Western; Cauliflower mosaic virus; Caulimovirus - genetics; DNA-Binding Proteins; Enzyme-Linked Immunosorbent Assay; Fundamental and applied biological sciences. Psychology; GAL4 promoter; gene expression; Genetic engineering; Genetic technics; genetic transformation; Genetic Vectors; Herpes simplex virus; Herpes Simplex Virus Protein Vmw65 - genetics; inducible promoter regions; Methods. Procedures. Technologies; Nicotiana - genetics; Nicotiana - virology; Nicotiana benthamiana; Plants, Genetically Modified - virology; promoter regions; Promoter Regions, Genetic; Proteins; recombinant DNA; reporter genes; Saccharomyces cerevisiae; Saccharomyces cerevisiae Proteins - genetics; TATA Box; Tobacco mosaic virus; Tobacco Mosaic Virus - genetics; transcription (genetics); Transcription Factors - genetics; Transcriptional Activation; transcriptional activator region; Transgenes; Transgenic animals and transgenic plants; transgenic plants; virus-mediated transgene activation; Virus; transgenic plants; Nicotiana benthamiana; GAL4; VP16; inducible promoters; Activation; Transgenic plant
• ISSN: 0962-8819; 1573-9368
• DOI: 10.1007/s11248-005-0388-7

We have developed a plant virus-mediated transgene activation (VMTA) system that utilizes a viral expression vector to present the inducer. The concept was tested using two well characterized components: (i) an artificial promoter based on the yeast GAL4 upstream activating sequence and the minimal TATA element of Cauliflower Mosaic Virus 35S RNA promoter, and (ii) a transcriptional activator (TA) consisting of a fusion between the GAL4 DNA binding domain and the Herpes simplex virus VP16 activation domain. The TA was expressed under the control of the subgenomic promoter of a Tobacco Mosaic Virus-based expression vector. The VMTA system was functional in transient Agroinfiltration assays with the reporter gene beta-glucuronidase, the intracellular domain of the diabetes associated autoimmune antigen, IA-2ic, and with the anti-tetanus antibody 9F12. Transgenic lines harboring the reporter gene were also examined. The VMTA system displayed tight transcriptional control in both transient assays and in transgenic Nicotiana benthamiana plants carrying the TA-inducible reporter.