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Development of Detection method for microcystins produced by toxic cyanobasteria

Research Project

Project/Area Number 08558056
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section展開研究
Research Field Environmental dynamic analysis
Research InstitutionIBARAKI UNIVERSITY

Principal Investigator

SHIRAI Makoto  IBARAKI Univ.Fuc.of Agriculture Prof., 農学部, 教授 (10007792)

Co-Investigator(Kenkyū-buntansha) SUZUKI Kouichi  Iatron Laboratories, Inc,, 研究所, 所長
KODAMA Osamu  IBARAKI UNIV.Fuc.of Agriculture Prof., 農学部, 教授 (00007791)
Project Period (FY) 1996 – 1998
Project Status Completed (Fiscal Year 1998)
Budget Amount *help
¥15,600,000 (Direct Cost: ¥15,600,000)
Fiscal Year 1998: ¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1997: ¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 1996: ¥10,100,000 (Direct Cost: ¥10,100,000)
Keywordsmicrocystin / waterbloom / hepaptotoxin / peptide toxin / antibody / fly / gene / cyanobacteria / ペプチド合成遺伝子 / 環状ペプチド / ラン藻 / Microcystis / Microcystis aeruginosa / モノクローナル抗体 / 毒素
Research Abstract

Toxic cyanobacterial (blue-green algal) waterblooms are found worldwide in eutrophic lakes and dams, and strains of several cyanobacterial genera produce the cyclic peptide hepatotoxin microcystins. The microcystins produced by cyanobacteria have become a serious environmental problem. In this studying, we developed three methods for detection of microcystins : (1) ELISA method, (2) Insecticidal bioassay method, (3) Gene probe and PCR method.
(1) ELISA method
Monoclonal antibodies against the microcystin LR variant were prepared from cloned hybridoma cell lines and ELISA method was developed.
(2) Insecticidal bioassay method
insecticidal bioassay for housefly was developed to investigate the toxicity of microcystin. The filter paper treated with microcystin in methanol was placed on a ethylene cup, then five adult houseflies were placed in the cup. After 24 hr, LD_<50> values of microcystin LR was 447 x 10^<-3> mu g/cm^2.
(3) Gene probe and PCR method
Microcystin synthetase genes were cloned and sequenced (ab. 45 kb). To amplify the microcystin synthetase genes, specific PCR primers were designed. Furthermore, specific DNA probes for detection of microcystin synthetase genes were designed. By using these probes and primers, microcystin-producing cells could be detected. Furthermore, regulation of gene expression in microcystin-producing Microcystis was revealed.

Report

(4 results)
  • 1998 Annual Research Report   Final Research Report Summary
  • 1997 Annual Research Report
  • 1996 Annual Research Report

Research Products

(37 results)

All Other

All Publications (37 results)

  • [Publications] M.Asayama: "Highly repetitivesequences and characteristics of genomic DNA in unicellular cyanobacterial strains." FEMS Mirobiol. Lett.137. 175-181 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] M.Asayama: "Cloning, sequencing and characterization of the gene encoding a principal sigma factor homolog from the cyanobacterium Microcystis aeruginosa K-81." Gene. 181. 213-217 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] M.Asayama: "The rpoD1 gene product is a Principal sigma factor of RNA polymerase in Microcystis aeruginosa K-81." J.Biochem.120. 752-758 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] 白井 誠: "アオコの毒性に関する研究" マイコトキシン. 42. 3-5 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] M.Sato: "Light-responsive and rhythmic gene expression of psbA2 in cyanobacterium Microcystis aeruginosa K-81." J.Gen.Appl.Microbiol.42. 381-391 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] I.Takahashi: "Restriction barrier composed of an extracellular nuclease and restriction endnuclease in the unicellular cyanobacterium Microcystis sp.." FEMS Microbiol. Lett.145. 107-111 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] 白井 誠: "富栄養化した湖沼に発生するアオコの毒性." モダンメディア. 42. 235-241 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] M.Asayama: "A new set of PCR primers for specific detection of the gene encoding the principal sigma factor in cyanobacteria." J.Gen.Appl.Microbiol.42. 511-515 (51)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] M.Asayama: "A novel genetic organization : the leuA-rpoD1 locus in the cyanobacterium Microcystis aeruginosa K-81." Biochim.Biopys.Acta. 1350. 15-20 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] M.Asayama: "A new sigma factor homolog in a cyanobacterium : cloning, sequencing, and light-responsive transcripts of rpoD2 from Microcystis aeruginosa K-81." Biochim.Biopys.Acta. 1351. 31-36 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] G.K.Agrawal: "A novel bend of DNA CIT : changeable bending-center sites of an intrinsic curvature under temperature conditions." FEMS microbiol.Lett.147. 139-145 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] J.Shibato: "Specific recognition of the cyanobacterial psbA promoter by RNA polymerases containing principal sigma factors. Biochim.Biophys.Acta." Biochim.Biopys.Acta.1442. 296-303 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] G.K.Agrawal: "Light-dependent and rhythmic psbA transcripts in homologous/heterologous cyanobacterial cells. Biochem." Biophys.Res.Comm.255. 47-53 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] M.Asayama: "An intrinsic DNA curvature found in the cyanobacterium Microcystis aeruginosa K-81 affects the promoter activity of rpoD1 encoding a principal sigma factor." J.Biochem.125. 460-468 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] 白井 誠: "医学微生物学の最先端(アオコの毒性と毒素遺伝子)" 菜根出版, 7 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] M.Asayama: "Highly repetitivesequences and characteristics of genomic DNA in unicellular cyanobacterial strains." FEMS Mirobiol.Lett.137. 175-181 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] M.Asayama: "Cloning, sequencing and characterization of the gene encoding a principal sigma factor homolog from the cyanobacterium Microcystis aeruginosa K-81" Gene. 181. 213-217 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] M.Asayama: "The ropD1 gene product is a principal sigma factor of RNA polymerase in Microcystis aeruginosa K-81" J.Biochem. 120. 752-758 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] M.Shirai: "Toxicity of natural blooms and Microcystis species (Japanese)" 42 Sup.3-5 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] M.Sato: "Light-responsive and rhythmic gene expression of psbA2 in cyanobacterium Microcystis aeruginosa K-81" J.Gen.Appl.Microbiol.42. 381-391 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] I.Takahashi: "Restriction barrier composed of an extracellular nuclease and restriction endnuclease in the unicellular cyanobacterium Microcystis sp" FEMS microbiol.Lett. 145. 107-111 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] M.Shirai: "Toxicity of the waterblooms (AOKO) occured in eutrophic water. (Japanese)" Modern Media. 42. 235-241 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] M.Asayama: "A new set of PCR primers for specfic detection of the gene encoding the principal sigma factor in cyanobacteria." J.Gen.Appl.Microbiol.42. 511-515 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] M.Asayama: "A novel genetic organization : the leuA-rpoD1 locus in the cyanobacterium Microcystis aeruginosa K-81" Biochim.Biopys.Aota. 1350. 15-20 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] M.Asayama: "A new sigma factor homolog in a cyanobacterium cloning, sequencing, and light-responsive transcripts of rpoD2 from Microcystis aeruginosa K-81" Biochim.Biopys.Acta.1351. 31-36 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] G.K.Agrawal: "A novel bend of DNA CIT.changeable bending center sites of an intrinsic curvature under temperature conditions." FEMS microbiol.Lett.147. 139-145 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] J.Shibato: "Specific recognition of the cyanobacterial psbA promoter by RNA polymerases containing principal sigma factors." Biochim.Biophys.Acta.1442. 296-303 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] G.K.Agrawal: "Light-dependent and rhythmic psbA transcripts in homologous /heterologous cyanobacterial cells." Biochem.Biophys.Res.Comm. 255. 47-53 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] M.Asayama: "An intrinsic DNA curvature found in the cyanobacterium Microcystis aeruginosa K-81 affects the promoter activity of rpoD1 encoding a principal sigma factor." J.Biochem. 125. 460-468 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] J.Shibato et al.: "Specific recognition of the cyanobacterial psbA promoter by RNA polymerases containing principal sigma factors." Biochim.Biophys.Acta.1442. 296-303 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] G.K.Agrawal et al.: "Light-dependent and rhythmic psbA transcripts in homologous/heterologous cyanobacterial cells." Biochem.Biophys.Res.Comm.255. 47-53 (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] M.Asayama et al.: "An intrinsic DNA curvature found in the cyanobacterium Microcystis aeruginosa K-81 affects the promoter activity of rpoD1 encoding a principal sigma factor." J.Biochem.,. 125. 460-468 (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] M.Asayama: "A new sigma factor homolog in a cyanobacterium" Biochim.Biophys.Acta. 1351. 31-36 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 白井 誠: "医学微生物学の最先端" 菜根出版, 7 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 白井誠: "富栄養化した湖沼に発生するアオコの毒性" モダンメディア. 42・6. 235-241 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] M.Asayama: "Highly repetitivesequences and characteristics of genomic DNA in unicellular cyanobacterial strains." FEMS Microbiol.Lett.137・2/3. 175-181 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] I.Takahashi: "Restriction barrier composed of an extracellular nuclease and restriction endnuclease in the unicellular cyanobacterium Microcystis sp.." FEMS Microbiol.Lett.145・1. 107-111 (1996)

    • Related Report
      1996 Annual Research Report

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Published: 1996-03-31   Modified: 2016-04-21  

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