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

  • Deborah Vicuna Requesens
  • Maria Elena Gonzalez Romero
  • Shivakumar P. Devaiah
  • Yeun-Kyung Chang
  • Ashley Flory
  • Stephen Streatfield
  • Rebecca Ring
  • Cassie Phillips
  • Nathan C. Hood
  • Cyrus Dean Marbaniang
  • John A. Howard
  • Elizabeth E. HoodEmail author
Original Paper
  • 91 Downloads

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.

Keywords

Maize Endosperm promoters Rice globulin Rice glutelin Maize α zein Recombinant protein expression 

Notes

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.

Supplementary material

11248_2019_162_MOESM1_ESM.doc (1.3 mb)
Supplementary material 1 (DOC 1320 kb)

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Copyright information

© Springer Nature Switzerland AG 2019

Authors and Affiliations

  • Deborah Vicuna Requesens
    • 1
    • 6
  • Maria Elena Gonzalez Romero
    • 1
  • Shivakumar P. Devaiah
    • 1
    • 7
  • Yeun-Kyung Chang
    • 1
    • 5
  • Ashley Flory
    • 1
  • Stephen Streatfield
    • 2
  • Rebecca Ring
    • 1
  • Cassie Phillips
    • 1
  • Nathan C. Hood
    • 3
    • 8
  • Cyrus Dean Marbaniang
    • 1
  • John A. Howard
    • 4
  • Elizabeth E. Hood
    • 1
    Email author
  1. 1.Arkansas State University Biosciences Institute and College of AgricultureArkansas State UniversityState UniversityUSA
  2. 2.Fraunhofer USA Center for Molecular BiotechnologyNewarkUSA
  3. 3.Infinite Enzymes, LLCJonesboroUSA
  4. 4.Applied Biotechnology InstituteSan Luis ObispoUSA
  5. 5.University of CalgaryCalgaryCanada
  6. 6.Coriell Institute for Medical ResearchCamdenUSA
  7. 7.BioStrategies, LCJonesboroUSA
  8. 8.St. LouisUSA

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