The Plant Pathology Journal

The Korean Society of Plant Pathology (한국식물병리학회)
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A Fluorescence-based cDNA-AFLP Method for Identification of Differentially Expressed Genes

  • Park, Sook-Young (Department of Agricultural Biotechnology, Center for Agricultural Biomaterials, Center for Fungal Genetic Resources, and Center for Fungal Pathogenesis, Seoul National University) ;
  • Jwa, Nam-Soo (Department of Molecular Biology, Sejong University) ;
  • Chi, Myoung-Hwan (Department of Agricultural Biotechnology, Center for Agricultural Biomaterials, Center for Fungal Genetic Resources, and Center for Fungal Pathogenesis, Seoul National University) ;
  • Lee, Yong-Hwan (Department of Agricultural Biotechnology, Center for Agricultural Biomaterials, Center for Fungal Genetic Resources, and Center for Fungal Pathogenesis, Seoul National University)
  • Published : 2009.06.30
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Abstract

Identification of differently expressed genes under specific tissues and/or environments provides insights into the nature and underlying mechanisms of cellular processes. Although cDNA-AFLP (Amplified Fragment Length Polymorphism) is a powerful method for analyzing differentially expressed genes, its use has been limited to the requirement of radioactive isotope use and the difficulty of isolating the bands of interest from a gel. Here, we describe a modified method for cDNA-AFLP that uses a fluorescence dye for detection and isolation of bands directly from a small size polyacrylamide gel. This method involves three steps: (i) preparation of cDNA templates, (ii) PCR amplification and differential display, and (iii) identification of differentially expressed genes. To demonstrate its utility and efficiency, differentially expressed genes during vegetative growth and appressorial development of Magnaporthe oryzae were analyzed. This method could be applied to compare gene expression profiles in a diverse array of organisms.

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