Transformation of Tobacco Cultured Cell by Particle Bombardment: Expression of Phenylalanine Ammonia-lyase Gene in Transgenic Tobacco Cells.

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  • NAGAI Nobuo
    Department of Applied Biology, Faculty of Textile Science and Technology, Shinshu University
  • YANAGISAWA Kuninori
    Department of Applied Biology, Faculty of Textile Science and Technology, Shinshu University
  • MIZUNO Kazuko
    Department of Applied Biology, Faculty of Textile Science and Technology, Shinshu University
  • IMURA Hirofumi
    Department of Applied Biology, Faculty of Textile Science and Technology, Shinshu University
  • TAGUCHI Goro
    Department of Applied Biology, Faculty of Textile Science and Technology, Shinshu University
  • SHIMOSAKA Makoto
    Department of Applied Biology, Faculty of Textile Science and Technology, Shinshu University
  • SHIBATA Daisuke
    Mitsui Plant Biotechnology Research Institute
  • OKAZAKI Mitsuo
    Department of Applied Biology, Faculty of Textile Science and Technology, Shinshu University

Bibliographic Information

Other Title
  • パーティクルガンによるタバコ培養細胞への遺伝子導入: phenylalanine ammonia-lyase (PAL) cDNAの導入と遺伝子発現
  • パーティクルガンによるタバコ培養細胞への遺伝子導入--phenylalanine ammonia-lyase(PAL)cDNAの導入と遺伝子発現〔英文〕
  • パーティクル ガン ニヨル タバコ バイヨウ サイボウ エ ノ イデンシ ドウ

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Description

The plasmid DNA containing a NPTII as a selectable marker gene and a tobacco PAL cDNA combined with the CaMV 35S promoter and NOS terminator was transformed into tobacco cells by a particle bombardment device based on flowing helium. The most efficient transformation was achieved when the amount of DNA-coated tungsten particle was 0.2mg per projectile, the amount of plasmid DNA was 4μg/mg of tungsten particles, the helium pressure accelerated was 3kg/cm2, and the distance between the sample and the projectile was 15cm. Genetic analysis of kanamycin-resistant transformants obtained showed that the PAL cDNA construct integrated into the genome of tobacco cells. PAL activity of the transformant increased almost 4-fold and scopoletin content increased more than 2-fold as compared to nontransformed cells.

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