Genomic approaches to the discovery of promoters for sustained expression in cotton (Gossypium hirsutum L.) under field conditions : expression analysis in transgenic cotton and Arabidopsis of a Rubisco small subunit promoter identified using EST sequence analysis and cDNA microarrays

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Biotechnology requires robust and predictable expression of transgenes. Most commercial Genetically Modified (GM) crops contain the viral 35S promoter to drive insecticide and herbicide resistance genes. In cotton there have been reductions in efficacy of <i>Bacillus thuringiensis</i> toxin (Bt) expressing plants late in the season that have been attributed to reductions in promoter activity. We have used genomic approaches to identify cotton genes whose expression remains high during the season to find promoters that might better maintain expression of transgenes in the field. A cDNA library from young late season leaves was used to generate about 2000 ESTs. Clustering of ESTs was used to determine relative transcript abundance and identify the most highly-expressed genes. These were primarily photosynthetic and housekeeping genes and some metabolic genes. The ESTs were printed to a small cDNA microarray and probed with both early- and late-season leaf mRNAs. Absolute fluorescence levels were used to rank genes and confirm the EST abundance data. Candidate genes, including the small subunit of Rubisco (RbcS) were selected. An RbcS promoter (Genbank Accession DQ648074) was isolated and analysed in both Arabidopsis and cotton linked to a GUS reporter gene. Expression of the reporter gene was consistently high in green tissues throughout the life cycle of cotton in the glasshouse, and in the field. A number of other candidate promoters have been identified that may be useful in a variety of biotechnology applications.

収録刊行物

  • Plant biotechnology  

    Plant biotechnology 23(5), 437-450, 2006-12-01 

    Japanese Society for Plant Cell and Molecular Biology

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各種コード

  • NII論文ID(NAID)
    10021909329
  • NII書誌ID(NCID)
    AA11250821
  • 本文言語コード
    ENG
  • 資料種別
    ART
  • ISSN
    13424580
  • NDL 記事登録ID
    8571128
  • NDL 雑誌分類
    ZR3(科学技術--生物学--植物)
  • NDL 請求記号
    Z54-J126
  • データ提供元
    CJP書誌  NDL  J-STAGE 
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