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Use of the wound-inducible NtQPT2 promoter from Nicotiana tabacum for production of a plant-made vaccine

Biotechnology Letters volume 34, pages 1143–1150 (2012)Cite this article

Abstract

The wound-inducible quinolinate phosphoribosyl transferase promoter from Nicotiana tabacum (NtQPT2) was assessed for its capacity to produce B-subunit of the heat-labile toxin (LTB) from enterotoxigenic Escherichia coli in transgenic plant tissues. Comparisons were made with the widely used and constitutive Cauliflower Mosaic Virus 35S (CaMV35S) promoter. The NtQPT2 promoter produced somewhat lower average concentrations of LTB protein per unit weight of hairy root tissue but allowed better growth thereby producing similar or higher overall average yields of LTB per culture batch. Transgenic tobacco plants containing the NtQPT2-LTB construct contained LTB protein in roots but not leaves. Moreover, wounding NtQPT2-LTB transgenic plants, by removal of apices, resulted in an approximate 500% increase in LTB levels in roots when analysed several days later. CaMV35S-LTB transgenic plants contained LTB protein in leaves and roots but wounding made no difference to their LTB content.

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Acknowledgments

GD is grateful to Kathleen De Boer and Suzy Ryan for technical advice, laboratory support and their extensive secondary metabolite work with regards to QPT2 promoter. GD acknowledges receipt of a postgraduate support scholarship from Monash University. This work was supported by Australian Research Council (ARC) Linkage Grant LP0882311 involving AMW and JDH.

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Affiliations

  1. Department of Forest and Ecosystem Science; Melbourne School of Land and Environment, The University of Melbourne, Creswick, VIC, 3363, Australia

    John D. Hamill

  2. School of Biological Sciences, Monash University, Clayton, VIC, 3800, Australia

    Giorgio De Guzman, Amanda M. Walmsley & Diane E. Webster

  3. Department of Microbiology, Monash University, Clayton, VIC, 3800, Australia

    Diane E. Webster

Authors
  1. Giorgio De Guzman
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  2. Amanda M. Walmsley
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  3. Diane E. Webster
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  4. John D. Hamill
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Corresponding author

Correspondence to John D. Hamill.

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10529_2012_879_MOESM1_ESM.pptx

Supplementary Fig. 1 Mean LTB (A) and nicotine (B) concentrations (μg/g Dry Weight [DW]) in hairy root cultures containing either the CaMV35S-LTB or NtQPT2-LTB construct. Data for panel A represents analysis of all species (8 root lines treated in triplicate), 24 hours after treatment with 25 µM MeJa (dissolved in ethanol - right column) compared with an ethanol control (left column). Panel B represents nicotine from only N. tabacum cultures, as there was no detectable nicotine present in the cultures of the other species (De Guzman et al. 2011). All data represent means ± S.E.M (PPTX 70 kb)

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De Guzman, G., Walmsley, A.M., Webster, D.E. et al. Use of the wound-inducible NtQPT2 promoter from Nicotiana tabacum for production of a plant-made vaccine. Biotechnol Lett 34, 1143–1150 (2012). https://doi.org/10.1007/s10529-012-0879-6

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  • DOI: https://doi.org/10.1007/s10529-012-0879-6

Keywords

  • Hairy roots
  • Molecular farming
  • Plant-made vaccine
  • Quinolinate phosphoribosyl transferase
  • Recombinant protein
  • Transgenic plant