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

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概要

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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 Bacillus thuringiensis 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.

• 2006-12-01

著者

• Dennis Elizabeth

  Csiro Act Aus

• Constable Greg

  Csiro Plant Industry

• Llewellyn Danny

  Csiro Plant Ind. Act Aus

• AMARASINGHE B.

  CSIRO Plant Industry

• FAIVRE-NITSCHKE Emmanuelle

  CSIRO Plant Industry

• WU Yingru

  CSIRO Plant Industry

• UDALL Joshua

  Department of Ecology, Evolution and Organismal Biology, Bessey Hall, Iowa State University

• Udall Joshua

  Department Of Ecology Evolution And Organismal Biology Bessey Hall Iowa State University

関連論文

• 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
• Global Gene Expression Responses to Waterlogging in Roots and Leaves of Cotton (Gossypium hirsutum L.)
• The FLX Gene of Arabidopsis is Required for FRI-Dependent Activation of FLC Expression
• Post-Translational Modifications of the Endogenous and Transgenic FLC Protein in Arabidopsis thaliana
• Transcript Profiling During Fiber Development Identifies Pathways in Secondary Metabolism and Cell Wall Structure That May Contribute to Cotton Fiber Quality
• Expression Profiling Identifies Genes Expressed Early During Lint Fibre Initiation in Cotton
• ABA Regulates Apoplastic Sugar Transport and is a Potential Signal for Cold-Induced Pollen Sterility in Rice

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