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ANALYSES OF GENE STRUCTURE AND FUNCTION BY ELECTRONMICROSCOPY

Research Project

Project/Area Number 61440011
Research Category

Grant-in-Aid for General Scientific Research (A)

Allocation TypeSingle-year Grants
Research Field 植物保護
Research InstitutionHOKKAIDO UNIVERSITY

Principal Investigator

SHIKATA Eishiro  HOKKAIDO UNIV., FACULTY OF AGR., PROFESSOR, 農学部, 教授 (00001389)

Co-Investigator(Kenkyū-buntansha) SANO Teruo  HOKKAIDO UNIV., FACULTY OF AGR., ASSISTANT, 農学部, 助手 (30142699)
UEDA Ichiro  HOKKAIDO UNIV., FACULTY OF AGR., ASSOC. PROF., 農学部, 助教授 (10113523)
IIZUKA Toshihiko  HOKKAIDO UNIV., FACULTY OF AGR., PROFESSOR, 農学部, 教授 (50001441)
OGOSHI Akira  HOKKAIDO UNIV., FACULTY OF AGR., PROFESSOR, 農学部, 教授 (80109503)
KINOSHITA Toshiro  HOKKAIDO UNIV., FACULTY OF AGR., PROFESSOR, 農学部, 教授 (10001421)
中村 太郎  北海道大学, 農学部, 教授 (30001390)
Project Period (FY) 1986 – 1989
Project Status Completed (Fiscal Year 1989)
Budget Amount *help
¥28,000,000 (Direct Cost: ¥28,000,000)
Fiscal Year 1989: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1988: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1987: ¥6,000,000 (Direct Cost: ¥6,000,000)
Fiscal Year 1986: ¥20,000,000 (Direct Cost: ¥20,000,000)
KeywordsElectron microscopy / Nucleic acid / Gene
Research Abstract

1) Cytochrome C method for visualizing double-standed DNA, double-stranded RNA, and single-stranded RNA was developed. The nucleic acids were rotary shadowed with Pt-Pd. BAC method was also applied. The method was simple and rapid when compared with the Cytochrome C method.
2) All the double-stranded genome segments of rice dwarf, rice black-streaked, and rice ragged stunt viruses were examined under the electron microscope for sizing. The sizes were in good agreement with those determined by nucleotide sequencing.
3) Non-chromosomal genetic elements of Rhizoctonia, a plant pathogen, were examined under the microscope. Linear dsRNA molecules were detected.
4) Hop stunt viroid and viroid from plum were seen as a rigid rod under the microscope. However, they were seen as a circular molecule 90-100 nm in length under the denaturing condition.
5) Mitochondrial DNAs from normal(N) and male sterile(S) cytoplasms of sugar beet were investigated by electron microscopy. The DNA was composed of a heterogeneous population of circular molecules. Their contour lengths varied from 0.28-51 mum. N and S cytoplasms were shown to contain their own characteristic combinations of small circular DNA species with lengths between 0.28-0.6 mum. Chloroplast DNA was a circular molecule of 45 mum in length.
6) The native plasmids bearing the crystal protein gene of Bacillus thuringiensis were observed by electron microscopy. The size of several large plasmids from subsop. sotto, kurstaki HD-1, and aizawai were determined by computer analysis of micrographs. The plasmids were a circular molecule and its estimated size was about 7OKb.
7) Viroplasm was isolated from rice dwarf virus infected rice plants by differential centrifugation. Viroplasm was seen in 5,000g fraction.

Report

(5 results)
  • 1989 Annual Research Report   Final Research Report Summary
  • 1988 Annual Research Report
  • 1987 Annual Research Report
  • 1986 Annual Research Report
  • Research Products

    (16 results)

All Other

All Publications (16 results)

  • [Publications] I.Uyeda et al.: "Size of dsRNA genome segments of rice dwarf and rice ragged stunt Viruses determined by electron microscopy" J.Electron Microsc.30. 34-58 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] T.Iizuka et al.: "Electron microscopic observation of the plasmid DNA bearing insecticidal crystal protein gene in Bacillus thuringiensis" The J.of Sericultural Science of Japan. 48. 448-456 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Y.Kijima et al.: "Restriction fragment map of sugar beet(Beta vulgris L.)Chloroplast DNA" Plant Molecular Biology. 7. 201-205 (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] T.Mikami: "Heterogeneity of circular mitochondrial DNA molecules from sugar beet with normal and male sterile cytoplasms." Curr.Genet.10. 695-700 (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Y. Kishima, T. Mikami, T. Harada, K. Shinozaki, M. Sugiura, and T. Kinoshita.: "Restriction fragment map of sugar beet (Beta vulgaris L.) chloroplast DNA." Plant Molecular Biology. 7. 201-205 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] T. Mikami, T. Harada, and T. Kinoshita.: "Heterogeneity of circular mitochondrial DNA molecules from sugar beet with normal and male sterile cytoplasms." Curr. Genet., 10 695-700, 1986.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] T. Iizuka, M. Arakida, H. Kikuta, K. Isida, I. Uyeda, and E. Shikata.: "Electron microscopic observation of the plasmid DNA bearing insecticidal crystal protein gene in Bacillus thuringiensis." The J. of Sericultural Science of Japan, 58 448-456, 1989.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] I. Uyeda, S. Y. Lee, G. W. Leng, G. M. Ao, Z. P. Deng, X. S. Liang, K. Ohshima, and E. Shikata.: "Size of dsRNA genome segments of rice dwarf and rice ragged stunt viruses determined by electron microscopy." J. Electron microsc., 39 34-58, 1990.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] I.Uyeda et al.: "Size of dsRNA genome segments of rice dwarf and rice ragged stunt Viruses detarmined by electron microscopy" J.Electron Microsc.30. 34-58 (1990)

    • Related Report
      1989 Annual Research Report
  • [Publications] T.Iizuka et al.: "Electron microscopic observation of the plasmid DNA bearing insecticidal crystal protein gene in bacillus thuringiensis" The J.of Sericultural Science of Japan. 48. 448-456 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Uyeda,I.;Kudo,H.;Takahashi,T.;Sano,T.;Ohshima,K.;Matsumura,T.;Shikata,E.: J.gen.Virol.70. (1989)

    • Related Report
      1988 Annual Research Report
  • [Publications] 飯塚 敏彦: Journal of Electron Microscopy. (1988)

    • Related Report
      1987 Annual Research Report
  • [Publications] I. Uyeda: Journal of Virological Methods.

    • Related Report
      1987 Annual Research Report
  • [Publications] I.Uyeda et al.: J.of Electron Microscopy. 35. 98-99 (1986)

    • Related Report
      1986 Annual Research Report
  • [Publications] Y.Kishima et al.: Plant Molecular Biology. 7. 201-205 (1986)

    • Related Report
      1986 Annual Research Report
  • [Publications] T.Mikami et al.: Current Genetics. 10. 695-700 (1986)

    • Related Report
      1986 Annual Research Report

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Published: 1987-03-31   Modified: 2016-04-21  

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