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The maize α-zein promoter can be utilized as a strong inducer of cellulase enzyme expression in maize kernels

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Abstract

Expression of recombinant proteins in plants is a technology for producing vaccines, pharmaceuticals and industrial enzymes. For the past several years, we have produced recombinant proteins in maize kernels using only the embryo, primarily driving expression of foreign genes with the maize globulin-1 promoter. Although strong expression is obtained, these lines use only 10–12% of the seed tissue. If strong embryo expression could be combined with strong endosperm expression, much more recombinant protein could be recovered from a set amount of seed biomass. In this study, we tested three endosperm promoters for expression of a cellulase gene. Promoters tested were rice globulin and glutelin promoters and a maize 19 kDa α-zein promoter. The rice promoters were used in two tandem expression constructs as well. Although the rice promoters were active in producing stable amounts of cellulase, the α-zein promoter was by far the most effective: as much as 9% of total soluble protein was recovered from seed of several independent events and plants. One or two inserts were detected by Southern blot in several lines, indicating that copy number did not appear to be responsible for the differences in protein accumulation. Tissue print analysis indicated that expression was primarily in the endosperm.

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Acknowledgements

This work was supported by Department of Energy Grant #DE FG36 GO88025; Walmart Foundation, Walton Family Foundation, and Arkansas State University Biosciences Institute. We would also like to acknowledge the technical assistance of Erin Egelkrout.

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Author notes

  1. Yeun-Kyung Chang

    Present address: University of Calgary, Calgary, AB, Canada

  2. Deborah Vicuna Requesens

    Present address: Coriell Institute for Medical Research, Camden, NJ, USA

  3. Shivakumar P. Devaiah

    Present address: BioStrategies, LC, Jonesboro, AR, 72401, USA

  4. Nathan C. Hood

    Present address: , St. Louis, MO, USA

Authors and Affiliations

  1. Arkansas State University Biosciences Institute and College of Agriculture, Arkansas State University, State University, AR, 72467, USA

    Deborah Vicuna Requesens, Maria Elena Gonzalez Romero, Shivakumar P. Devaiah, Yeun-Kyung Chang, Ashley Flory, Rebecca Ring, Cassie Phillips, Cyrus Dean Marbaniang & Elizabeth E. Hood

  2. Fraunhofer USA Center for Molecular Biotechnology, Newark, DE, 19711, USA

    Stephen Streatfield

  3. Infinite Enzymes, LLC, Jonesboro, AR, 72401, USA

    Nathan C. Hood

  4. Applied Biotechnology Institute, San Luis Obispo, CA, 93407, USA

    John A. Howard

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  1. Deborah Vicuna Requesens
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Correspondence to Elizabeth E. Hood.

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Vicuna Requesens, D., Gonzalez Romero, M.E., Devaiah, S.P. et al. The maize α-zein promoter can be utilized as a strong inducer of cellulase enzyme expression in maize kernels. Transgenic Res 28, 537–547 (2019). https://doi.org/10.1007/s11248-019-00162-1

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