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Transgenic Research

, Volume 14, Issue 5, pp 583–592 | Cite as

Improvement of Human lysozyme Expression in Transgenic Rice Grain by Combining Wheat (Triticum aestivum) puroindoline b and Rice (Oryza sativa) Gt1 Promoters and Signal Peptides

  • Kevin Hennegan
  • Daichang YangEmail author
  • Diane Nguyen
  • Liying Wu
  • Jeff Goding
  • Jianmin Huang
  • Fengli Guo
  • Ning Huang
  • Simon C. Watkins
Article

Abstract

Heterologous protein expression levels in transgenic plants are of critical importance in the production of plant-made pharmaceuticals (PMPs). We studied a puroindoline b promoter and signal peptide (Tapur) driving human lysozyme expression in rice endosperm. The results demonstrated that human lysozyme expressed under the control of the Tapur cassette is seed-specific, readily extractable, active, and properly processed. Immuno-electron microscopy indicated that lysozyme expressed from this cassette is localized in protein bodies I and II in rice endosperm cells, demonstrating that this non-storage promoter and signal peptide can be used for targeting human lysozyme to rice protein bodies. We successfully employed a strategy to improve the expression of human lysozyme in transgenic rice grain by combining the Tapur cassette with our well established Gt1 expression system. The results demonstrated that when the two expression cassettes were combined, the expression level of human lysozyme increased from 5.24 ± 0.34 mg−1 g flour for the best single cassette line to 9.24 ± 0.06 mg−1 g flour in the best double cassette line, indicating an additive effect on expression of human lysozyme in rice grain.

Keywords

gene stacking human lysozyme protein bodies puroindoline b transgenic rice 

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Copyright information

© Springer 2005

Authors and Affiliations

  • Kevin Hennegan
    • 1
  • Daichang Yang
    • 1
    Email author
  • Diane Nguyen
    • 1
  • Liying Wu
    • 1
    • 3
  • Jeff Goding
    • 1
  • Jianmin Huang
    • 1
  • Fengli Guo
    • 2
  • Ning Huang
    • 1
  • Simon C. Watkins
    • 2
  1. 1.Ventria BioscienceSacramentoUSA
  2. 2.Department of Cell Biology and PhysiologyUniversity of PittsburghUSA
  3. 3.Arcadia BioscienceDavisUSA

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