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


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Article Info

DATE

2008-02

AUTHORS

Maria Oszvald, Mark Gardonyi, Cecília Tamas, Imre Takacs, Barnabas Jenes, Laszlo Tamas

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. More... »

PAGES

1-7

References to SciGraph publications

  • 1998-03. Modifying transient β-glucuronidase expression in pine species using introns in PLANT CELL, TISSUE AND ORGAN CULTURE (PCTOC)
  • 2007-05. Promoter analysis and immunolocalisation show that puroindoline genes are exclusively expressed in starchy endosperm cells of wheat grain in PLANT MOLECULAR BIOLOGY
  • 1989-05. The characterization and comparative analysis of high-molecular-weight glutenin genes from genomes A and B of a hexaploid bread wheat in THEORETICAL AND APPLIED GENETICS
  • 1993-01. Analysis of a genomic DNA segment carrying the wheat high-molecular-weight (HMW) glutenin Bx17 subunit and its use as an RFLP marker in THEORETICAL AND APPLIED GENETICS
  • 2004-02. Stable transformation of rice (Oryza sativa L.) via microprojectile bombardment of highly regenerative, green tissues derived from mature seed in PLANT CELL REPORTS
  • 1997-12. Context sequences of translation initiation codon in plants in PLANT MOLECULAR BIOLOGY
  • 1991-01. Intron enhancement of gene expression and the splicing efficiency of introns in maize cells in MOLECULAR GENETICS AND GENOMICS
  • 1996-05. Ubiquitin promoter-based vectors for high-level expression of selectable and/or screenable marker genes in monocotyledonous plants in TRANSGENIC RESEARCH
  • 2007-03. Expression of a synthetic neutralizing epitope of porcine epidemic diarrhea virus fused with synthetic B subunit of Escherichia coli heat labile enterotoxin in rice endosperm in MOLECULAR BIOTECHNOLOGY
  • 2007-02. Somaclonal variation and stability of GUS gene expression in transgenic agapanthus (Agapanthus praecox ssp. orientalis) plants at the flowering stage in IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY - PLANT
  • 1991-12. Construction of expression vectors based on the rice actin 1 (Act1) 5′ region for use in monocot transformation in MOLECULAR GENETICS AND GENOMICS
  • 2004-11. Dissemination of the highly expressed Bx7 glutenin subunit (Glu-B1al allele) in wheat as revealed by novel PCR markers and RP-HPLC in THEORETICAL AND APPLIED GENETICS
  • 2003-08. Bánkúti 1201—an old Hungarian wheat variety with special storage protein composition in THEORETICAL AND APPLIED GENETICS
  • 2001-03. Designing of an artificial expression cassette for the high-level expression of transgenes in plants in THEORETICAL AND APPLIED GENETICS
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    http://scigraph.springernature.com/pub.10.1007/s11627-007-9082-1

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    http://dx.doi.org/10.1007/s11627-007-9082-1

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    32 schema:description 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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