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

Oszvald, Mária and Gardonyi, M. and Tamás, Cecília and Takács, Imre and Tamás, László (2008) Development and characterization of a chimaeric tissue-specific promoter in wheat and rice endosperm. IN VITRO CELLULAR AND DEVELOPMENTAL BIOLOGY-PLANT, 44 (1). pp. 1-7. ISSN 1054-5476

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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.

Item Type: Article
Subjects: Q Science / természettudomány > QH Natural history / természetrajz > QH301 Biology / biológia
Q Science / természettudomány > QK Botany / növénytan
SWORD Depositor: MTMT SWORD
Depositing User: MTMT SWORD
Date Deposited: 23 May 2013 09:06
Last Modified: 23 May 2013 09:06
URI: http://real.mtak.hu/id/eprint/5227

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