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Inducible expression in plants by virus-mediated transgene activation

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Abstract

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 theHerpes 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 β-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.

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Abbreviations

dpi:

days post inoculation

GFPC3:

green fluorescent protein cycle 3

GUS:

β-glucuronidase

IA-2ic:

intracellular domain of islet antigen

TA:

transcriptional activator

TMV:

Tobacco Mosaic Virus

VMTA:

virus mediated transgene activation

VP16:

herpes simplex viral protein 16

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Authors and Affiliations

  1. Fraunhofer USA Center for Molecular Biotechnology, 9 Innovation Way, Suite 200, Newark, Delaware, 19711, USA

    Anna K. Hull, Vidadi Yusibov & Vadim Mett

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  1. Anna K. Hull
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  2. Vidadi Yusibov
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  3. Vadim Mett
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Correspondence to Vadim Mett.

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Hull, A.K., Yusibov, V. & Mett, V. Inducible expression in plants by virus-mediated transgene activation. Transgenic Res 14, 407–416 (2005). https://doi.org/10.1007/s11248-005-0388-7

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