Construction of expression vectors based on the rice actin 1 (Act1) 5′ region for use in monocot transformation View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

1991-12

AUTHORS

David McElroy, Alan D. Blowers, Barnabás Jenes, Ray Wu

ABSTRACT

It has been previously reported that the 5' region of the rice actin 1 gene (Act1) promoted high-level expression of a beta-glucuronidase reporter gene (Gus) in transformed rice cells. In this paper we describe the construction of Act1-based expression vectors for use in monocot transformation. As part of the development of these vectors, we have evaluated the influence of the Act1 first intron, the Act1-Gus junction-encoded N-terminal amino acids, and the sequence context surrounding the Act1 and Gus translation initiation site on Act1-Gus gene expression in rice and maize cells. We have found that addition of Act1 intron 1 to the transcription unit of a Gus reporter gene under control of the cauliflower mosaic virus (CaMV) 35S promoter stimulated GUS activity more than 10-fold in transformed rice cells. Optimization of the sequence context around the Gus translation initiation site resulted in a 4-fold stimulation of Gus expression in transformed rice cells. By utilizing both the Act1 intron 1 and optimized Gus translation initiation site, a 40-fold stimulation in Gus expression from the CaMV 35S promoter has been achieved in transformed rice cells; very similar results were obtained in transformed maize cells. Taken together these results suggest that the Act1-based expression vectors described here should promote the expression of foreign genes in most, if not all, transformed monocot cells to levels that have not previously been attainable with alternative expression vectors. More... »

PAGES

150-160

References to SciGraph publications

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  • 1990-12. Intron-mediated enhancement of heterologous gene expression in maize in PLANT MOLECULAR BIOLOGY
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  • 1989-03. Chimeric gene expression using maize intron in cultured cells of breadwheat in PLANT CELL REPORTS
  • 1990-07. Recognition efficiency ofDicotyledoneae-specific promoter and RNA processing signals in rice in MOLECULAR GENETICS AND GENOMICS
  • 1987-12. Assaying chimeric genes in plants: The GUS gene fusion system in PLANT MOLECULAR BIOLOGY REPORTER
  • 1988-12. Efficient regeneration of transgenic plants from rice protoplasts and correctly regulated expression of the foreign gene in the plants in THEORETICAL AND APPLIED GENETICS
  • 1991-01. Intron enhancement of gene expression and the splicing efficiency of introns in maize cells in MOLECULAR GENETICS AND GENOMICS
  • 1990-05. Bialaphos selection of stable transformants from maize cell culture in THEORETICAL AND APPLIED GENETICS
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  • 1991-02. The combination of a novel stimulatory element in the first exon of the maize Shrunken-1 gene with the following intron 1 enhances reporter gene expression up to 1000-fold in PLANT MOLECULAR BIOLOGY
  • 1990-02. Structural characterization of a rice actin gene in PLANT MOLECULAR BIOLOGY
  • 1988-05. Factors Influencing Gene Delivery into Zea Mays Cells by High–Velocity Microprojectiles in NATURE BIOTECHNOLOGY
  • 1990-02. Expression of CaMV35S-GUS gene in transgenic rice plants in MOLECULAR GENETICS AND GENOMICS
  • 1991-08. Anaerobic induction and tissue-specific expression of maize Adh1 promoter in transgenic rice plants and their progeny in MOLECULAR GENETICS AND GENOMICS
  • Journal

    TITLE

    Molecular Genetics and Genomics

    ISSUE

    1

    VOLUME

    231

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  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1007/bf00293832

    DOI

    http://dx.doi.org/10.1007/bf00293832

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1050086821

    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/1753941


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