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Development and characterization of a chimaeric tissue-specific promoter in wheat and rice endosperm

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Abstract

The recently achieved significant improvement of cereal transformation protocols provides facilities to alter the protein composition of the endosperm, for example, to increase or decrease the quantity of one of its protein components or to express foreign molecules. To achieve this goal, strong endosperm-specific promoters have to be available. The aim of our work was to develop a more efficient tissue-specific promoter which is currently used. A chimaeric promoter was assembled using the 5′ UTR (1,900 bp) of the gene coding for the 1Bx17 HMW glutenin subunit protein, responsible for tissue-specific expression and the first intron of the rice actin gene (act1). The sequence around of the translation initial codon was optimized. The effect of the intron and promoter regulatory sequences, using different lengths of 1Bx17 HMW-GS promoter, were studied on the expression of uidA gene. The function of promoter elements, promoter length, and the first intron of the rice actin gene were tested by a transient expression assay in immature wheat endosperm and in stable transgenic rice plants. Results showed that insertion of the rice act1 first intron increased GUS expression by four times in transient assay. The shortest 1Bx17 HMW-GS promoter fragment (173 bp) linked to the intron and GUS reporter gene provided almost the same expression level than the intronless long 1Bx17 HMW-GS promoter. Analysis of the stable transformant plants revealed that 173 nucleotides were sufficient for endosperm-specific expression of the uidA gene, despite 13 nucleotides missing from the HMW enhancer sequence, a relevant regulatory element in the promoter region.

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Acknowledgments

This work was supported by the Hungarian Scientific Research Fund (OTKA T 034791) and partly by ICGEB (CRP/HUN00-02).

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Authors and Affiliations

  1. Department of Biochemistry and Food Technology, BUTE University, Budapest, Hungary

    Maria Oszvald

  2. Department of Plant Physiology and Molecular Plant Biology, Eotvos Lorand University, Pázmány P. sétány 1/C., 1117, Budapest, Hungary

    Mark Gardonyi & Laszlo Tamas

  3. Agricultural Biotechnology Center, Gödöllő, Hungary

    Maria Oszvald & Barnabas Jenes

  4. Agricultural Research Institute of the HAS, Martonvásár, Hungary

    Cecília Tamas & Imre Takacs

Authors
  1. Maria Oszvald
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  2. Mark Gardonyi
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  3. Cecília Tamas
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  4. Imre Takacs
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  5. Barnabas Jenes
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  6. Laszlo Tamas
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Correspondence to Laszlo Tamas.

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K. D’Halluin

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Oszvald, M., Gardonyi, M., Tamas, C. et al. Development and characterization of a chimaeric tissue-specific promoter in wheat and rice endosperm. In Vitro Cell.Dev.Biol.-Plant 44, 1–7 (2008). https://doi.org/10.1007/s11627-007-9082-1

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  • DOI: https://doi.org/10.1007/s11627-007-9082-1

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