Journal of Photoscience

Korean Society of Photoscience (한국광과학회)
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Random Insertional Mutagenesis with Subtracted cDNA Fragments in Arabidopsis thaliana

  • Euna Cho (School of Biological Sciences, Seoul National University) ;
  • Kwon, Young-Myung (School of Biological Sciences, Seoul National University) ;
  • Lee, Ilha (Plant metabolosm ResearchCenter, Kyung Hee University)
  • Published : 2000.09.01
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Abstract

We have evaluated a new mutagenesis strategy called random insertional mutagenesis with subtracted cDNA fragments. The cDNAs from long day Arabidopsis plants were subtracted by cDNAs from short day plants using PCR based cDNA subtraction. The subtracted cDNAs were inserted between 35S promoter and 3'-NOS terminator regardless of orientation. When the cDNA library was used for the random insertion into Arabidopsis genome by Agrobacterium-mediated transformation, approximately 15% of transformants showed abnormal development in leaf, floral organ, shoot apex. When 20 mutants were analyzed, 12 mutants showed single cDNA fragment insertion and 8 mutants showed more than 2 transgene insertions. Only two mutants among 12 mutants that have single cDNA insert showed consistent phenotype at T2 generation, suggesting the genetic instability of the mutants.

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