Isolation and analysis of CPD photolyase gene of spinach and fundamental study for the production pf transgenic plant resistant to UV

• Project/Area Number: 13680628
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: 環境影響評価(含放射線生物学)
• Research Institution: Yamaguchi University
• Principal Investigator: TAKIMOTO Koichi Faculty of Agriculture, professor, 農学部, 教授 (00115875)
• Co-Investigator(Kenkyū-buntansha): MANO Junichi Faculty of Agriculture, associate professor, 農学部, 助教授 (50243100) ; MURAKAMI Ryutaro Faculty of Science, associate professor, 理学部, 助教授 (40182109)
• Project Period (FY): 2001 – 2002
• Project Status: Completed (Fiscal Year 2002)
• Budget Amount: ¥3,600,000 (Direct Cost: ¥3,600,000); Fiscal Year 2002: ¥900,000 (Direct Cost: ¥900,000); Fiscal Year 2001: ¥2,700,000 (Direct Cost: ¥2,700,000)
• Keywords: ultraviolet light / DNA damage / DNA repair / pvrimidine dimers / photoreactivation / photoreactivating enzyme / spinach / plant / ピリミジン二量体

Research Abstract

We isolated a gene for cyclobutane pyrimidine dimer (CPD) photolyase in spinach and determined the function and the extent of the expression in male and female plants. cDNA of the gene was cloned by RTPCR with the use of niRNA as a tremplate and degenerate primers. The gene consists of nine exsons and amino acid sequence was quite similar to class II CPD photolyase. The gene obtined here restored the photoreactivating ability of mutant E. coil deficient in photorepair indicating that the gene was photolyase gene. Removal of CPD was confirmed by ELISA procedure. The gene was a single copy. The level of expression in root was lower than in leaves, stems and flowers in male and female spinach. UV generates reactive oxygen species (ROS). Specificity of ROS mutation was investigated in E. coil. Base change mutations were predominant among mutations.

Research Products

• [Publications] Tano, K., Iwamatsu, Y., Utsumi, H., Takimoto, K.: "Increased base change mutations at G:C pairs in Escherichia coli deficient in endonuclease III and VIII"J. Radiat. Res.. 42・4. 409-413 (2001)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Mano, J., Torii, Y., Hayashi, S., Takimoto, K., Matsui, K., Nakamura, K., Inze, D., Babiychuk, E., Kushnir, S., Asada, K.: "The NADPH : Quinone oxidoreductase P1-ζ-crystallin in Arabidopsis catalyzes the a,b-hydrogenation of 2-alkenals : Detoxication of the lipid peroxide-derived reactive aldehydes"Plant Cell Physiol.. 43・12. 1445-1455 (2002)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Tano, K., Iwamatsu, Y., Utsumi, H. & Takimoto, K.: "Increased base change mutations at G:C pairs in Escherichia coli"J. Radiat. Res.. 42(4). 409-413 (2001)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Mano J., Toni, Y., Hayashi, S., Takimoto, K., Matsui, K., Nalcam Plant Cell Physiol. 43(12) 1445-1455 tira, IC, line, D., Babiychuk, K, Kushnir, S. & Asada, K.: "The NADPH: Quinone oxidoreductase P1-C-crystallin in Arabidopsis catalyzes the α,β-hydrogenation of 2-alkenals: Detoxication of the lipid peroxide-derived reactive aldehydes"Plant Cell Physiol. 43(12). 1445-1455 (2002)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Keizo Tano: "Increased base change mutations at G: C pairs in Escherichia coil deficient in endonuclease III and VIII"J. Radiat. Res.. 42.4. 409-413 (2001)
• [Publications] Junichi Mano: "The NADPH: Quinone oxidoreductase P1-ζ-crystallin in Arabidopsis catalyzes the a, b-hydrogenation of 2-alkenals : Detoxication of the lipid peroxide-derived reactive aldehydes"Plant Cell Physiol.. 43.12. 1445-1455 (2002)
• [Publications] K.Tano, Y.Iwamatsu, S.Yasuhira, Utsumi, H., K.Takimoto: "Increased base change mutations at G : C pairs in Escherichia coli deficient in Endonuclease III and VIII"J. Radiat Res.. 42(4). 409-413 (2001)