• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to project page

1994 Fiscal Year Final Research Report Summary

Cloning and Structural Analysis of Na^+-Translocating NADH-Quinone Reductase of Marine Bacteria

Research Project

Project/Area Number 05671809
Research Category

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

Allocation TypeSingle-year Grants
Research Field Biological pharmacy
Research InstitutionChiba University

Principal Investigator

UNEMOTO Tsutomu  Chiba University, Faculty of Pharm. Sciences Professor, 薬学部, 教授 (30089601)

Co-Investigator(Kenkyū-buntansha) HAYASHI Maki  Chiba University, Faculty of Pharm. Sciences Associate Professor, 薬学部, 助教授 (50092086)
Project Period (FY) 1993 – 1994
KeywordsSodium ion transport / Sidium pump / NADH-quinone reductase / Cloning of NQR gene / Respiratory chain / Marine bacteria / Vibrio alginolyticus / Vibrio alginolyticus
Research Abstract

The Na^+-translocating NADH-quinone reductase purified from the marine bacterium Vibrio alginolyticus is composed of three subunits, alpha, beta, and gamma. Each subunit was purified by SDS-polyacryl-amide gel electrophoresis and each N-terminal amino acid sequence was determined. Each subunit was also partially digested with V8 protease, and the N-terminal amino acid sequence of the purified polypeptide fragment was determined. From these informations, oligonucleotides corresponding to forward and reverse primers for each gene (NQR A,B and C) encoding alpha, beta and gamma subunits, respectively, were synthesized. Using these primers, a part of each gene was amplified from the chromosomal DNA of V.alginolyticus by a PCR method, and the PCR products were used for the cloning of NQR gene in lambda phage. Among the subclones selected by the probe C,the expression beta-subunit as a gene product was detected in Escherichia coli membranes by the activity staining and by Western blotting analyzes.

  • Research Products

    (6 results)

All Other

All Publications (6 results)

  • [Publications] Unemoto,T.: "Na^+Translocating NADH-Quinone Reductase of Marine and Halophilic Bacteria." Journal of Bioenergetics and Biomembranes,. 25. 385-391 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Unemoto,T.: "Modifications by Na^+ and K^+,and the Site of Ag^+Inhibition in the Na^+ Translocating NADH-Quinone Reductase from a Marine Vibrio alginolyticus." Biochimica et Biophysica Acta.1183. 201-205 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hayashi,M.: "Cloning of Na^+-Translocating NADH-Quinone Reductase Gene from the Marine Vibrio alginolyticus and the Expression of β-Subunit in Escherichia coli." FEBS Letters,. 356. 330-332 (1994)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Tsutomu Unemoto and Maki Hayashi: "Na^+-Translocating NADH-Quinone Reductase of Marine and Halophilic Bacteria." Journal of Bioenergetics and Biomembranes. Vol.25, No.4. 385-391 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Tsutomu Unemoto, Tatsuya Ogura and Maki Hayashi: "Modifications by Na^+ and K^+, and the Site of Ag^+ Inhibition in the Na^+-Translocating NADH-Quinone Reductase from a Marine Vibrio alginolyticus." Biochmica et Biophysica Acta. Vol.1183. 201-205 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Maki Hayashi, Kouji Hirai and Tsutomu Unemoto: "Cloning of Na^+-Translocating NADH-Quinone Reductase Gene from the Marine Vibrio alginolyticus and the Expression of beta-Subunit in Escherichia coli." FEBS Letters. Vol.356. 330-332 (1994)

    • Description
      「研究成果報告書概要(欧文)」より

URL: 

Published: 1996-04-15  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi