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Construction of transgenic mosquitoes for foreign gene expression

Research Project

Project/Area Number 11680827
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Laboratory animal science
Research InstitutionOITA MEDICAL UNIVERSITY

Principal Investigator

ESHITA Yuki  Oita Medical University, Faculty of Medicine, Associate Professor, 医学部, 助教授 (10082223)

Co-Investigator(Kenkyū-buntansha) FUKUDA Masako  Oita Medical University, Faculty of Medicine, Research Associate, 医学部, 助手 (00156788)
MATSUMOTO Akira  Kyushu University Research Center for Higher Education, Research Associate, 大学教育研究センター, 助手 (40229539)
Project Period (FY) 1999 – 2001
Project Status Completed (Fiscal Year 2001)
Budget Amount *help
¥3,700,000 (Direct Cost: ¥3,700,000)
Fiscal Year 2001: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 2000: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1999: ¥1,500,000 (Direct Cost: ¥1,500,000)
Keywordspathogen resistant mosquitoes / mariner genome / injection and electroporation methods / IE 1 promoter / GFP / seamless cloning method / SV40 / baculovirus / 日本産蚊 / mariner / transposase / 転写終結シグナル / actin5C / EGFP / トランスポゾン / 組換え蚊 / 電気窄孔法 / 注入法 / ホーボ遺伝子 / 反復逆配列 / 外来遺伝子の発現
Research Abstract

Using the genetic engineering techniques, the procedure for producing the pathogen-resistant mosquitoes in Japan was examined.
1. Existence of the mariner-like gene in Japanese mosquito genomes.
Inversed terminal repeat (ITR) of the mariner-like genes as a primer was used for the amplification of the mariner overall length. As the results, there was the amplification of the ITR gene in not only Aedes albopictus mosquitoes but also Anopheles species. However, the PCR product was observed only in the Aedes albopictus mosquitoes, when conserved region in the transferase enzyme coded in the mariner gene was used as a primer. From these observations, it may be suggested that mariner-like gene functions in the Aedes albopictus mosquito as one of active transposons.
2. Hatchability of mosquito eggs post injection.
Mixture DNAs of the hobo gene that is a kind of the transposon, and of transposase as the helper were injected into eggs of Aedes aegypti post 90 minutes progress. And afterwards, their … More hatchability were observed. As the results, 39 % of Aedes aegypti eggs injected hatched as normal larvae. However, no Aedes albopictus eggs injected with DNA solutions hatched.
It was indicated that injection condition to the mosquito egg differed by mosquito species.
3. Hatchability of mosquito eggs post electroporation.
As a method for inserting mariner [actin5C-EGFP] gene into mosquito genomes, injection and electroporation methods were examined. Mariner [actin5C-EGFP] DNA as an artificial transposon and helper (mariner) DNA mixture were used. Aedes aegypti eggs hatched by electroporation method using pulsed apparatus. However, no hatched egg of Aedes albopictus was observed. It was suggested that the suitable pulse condition differed in the mosquito species.
4. Construction of an artificial transfer vector using an active transposon derived from Japanese mosquitoes
Enhancer and IE 1 promoter derived from baculoviruses in the upperstream of an active transposon mariner element of Japanese mosquitoes, and the GFP gene as a marker in the downstream were constructed. More details were described in this report. Less

Report

(4 results)
  • 2001 Annual Research Report   Final Research Report Summary
  • 2000 Annual Research Report
  • 1999 Annual Research Report
  • Research Products

    (27 results)

All Other

All Publications (27 results)

  • [Publications] Eshita, Y.: "Vector competence of Japanese mosquitoes against dengue viruses"SP World. No.29. 13-17 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Uchida, K., Ohmori, D., Ueno, T., Nishizuka, M., Eshita Y., et al.: "Preoviposition activation of cathepsin-like proteinases in degenerating ovarian follicles of the mosquito Culex pipiens pallens"Developmental Biology. 237. 68-78 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Oda, T., Eshita, Y., Uchida, K., Mine, M., Kurokawa, K., et al.: "Reproductive activity and survival of Culex pipiens pallens and Culex quinquefasciatus (Diptera: Culicidae) in Japan at high temperature"J.Med.Entomol.. 39. 185-190 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Morales, R., Morita, K., Eshita, Y., Tsuda, Y., Fukuma, T., Takagi, M.: "Infection and dissemination of two dengue type 2 viruses isolated from patients exhibiting different disease seventies in orally infected Aedes aegypti from different geographic origin"Med.Entomol.Zool.. 53(in print). (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Eshita, Y., Fukuda, M., Anzal, S., Otsuka, Y., Aoki, C., Takaoka, H., et al.: "Biologia y epidemiologia molecular del mosquito vector del virus del dengue [in Spanish]"Cemadoja Cientifica (Republica Dominicana). 2(in print). (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Eshita, Y., Fukuda, M., Anzai, S., Takaoka, H.Igarashi, A., et al.: "Competencia del mosquito como vector del virus del dengue[in Spanish]"Cemadoja Cientifica (Republica Dominicana). 2(in print). (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] 江下優樹, 松本顕: "地球規模での寄生虫対策の時代(分担執筆)別冊・医学のあゆみ"医歯薬出版株式会社. 132 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Eshita, Y.: "Vector competence of Japanese mosquitoes against dengue viruses"SP World. No.29. 13-17 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Uchida, K., Ohmori, D., Ueno, T., Nishizuka, M., Eshita Y., Fukunaga, A. and Kominami E.: "Preoviposition activation of cathepsin-like proteinases in degenerating ovarian follicles of the mosquito Culex pipiens pallens"Developmental Biol.. 237. 68-78 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Oda, T., Eshita, Y., Uchida, K., Mine, M., Kurokawa, K., Ogawa, Y., Kato, K. and Tahara, H.: "Reproductive activity and survival of Culex pipiens pattern and Culex quinquefasciatus (Diptera : Culicidae) in Japan at high temperature"J. Med. Entomol.. 39(1). 185-190 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Morales, R., Morita, K., Eshita, Y., Tsuda, Y., Fukuma, T., and Takagi, M.: "Infection and dissemination of two dengue type 2 viruses isolated from patients exhibiting different disease severities in orally infected Aedes aegypti from different geographic origin"Med. Entomol. Zool.. (in print).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Eshita, Y., Fukuda, M., Anzai, S., Otsuka, Y., Aoki, C., Takaoka, H., Matsumoto, A., Uchida, K., Igarashi, A., Uchida, Y., Takasaki, T., Yamada, K., y Kurane I.: "Biologia y epidemiologia molecular del mosquito vector del virus del vengue [in Spanish]"Cemadoja Cientifica (Republica Dominicana). (in print).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Eshita, Y., Fukuda, M., Anzai, S., Takaoka. H., Igarashi, A., Uchida, Y., Takasaki, T., Yamada, K., y Kurane I.:: "Competencia del mosquito como vector del virus del dengue [in Spanish]"Cemadoja Cientifica (Republica Dominicana). (in print).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Eshita, Y., and Matsumoto, A: "Control of vector insects - Development of pathogen-resistant mosquitoes using genetic engineering (in Japanese)"In : The age of parasitic disease control at the global scale, (Editor : Tada, I.), pp98 -103, Ishiyaku Publishers, Inc., Tokyo. (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] 江下優樹, 松本 顕: "伝播昆虫の制御:遣伝子工学を用いた病原体耐性蚊の開発"医学のあゆみ. 191・1. 98-103 (1999)

    • Related Report
      2001 Annual Research Report
  • [Publications] Eshita, Y.: "Vector competence of Japanese mosquitoes against dengue viruses"SP World. No.29. 13-17 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Takaoka, H..Bain, O., UnI, S., Korenaga, M., Tada.K..Ichikawa, H., Otsuka.Y., Eshita.Y.: "Human infection with Onchocerca dewittei japonica, a parasite from wild boar in Oita, Japan"Parasite. 8・3. 261-263 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Uchida, K., Oda T., Matsuoka, H., Moribayashi, A., Ohmori, D., Eshita Y.: "Induction of oogenesis in mosquitoes (Diptera : Culicidae) by Infusion of the hemocoel with amino acids"J.Med.Entomol.. 38・4. 572-575 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Uchida, K., Ohmori, D., Ueuo, T., Nishizuka, M., Eshita Y., et al.: "Preoviposition activation of cathepsin-like proteinases in degenerating ovarian follicles of the mosquito Culex piplens pallens"Developmental Biology. 237. 68-78 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Oda, T., Eshita, Y., Uchida, K., Mine, M., Kurokawa, K., et al.: "Reproductive activity and survival of Culex pipiens pallens and Culex quinquefasciatus (Diptera : Culicidae) in Japan at high temperature"J.Med.Entomol.. 39・1. 185-190 (2002)

    • Related Report
      2001 Annual Research Report
  • [Publications] Oda,T.,Uchida,K.,Mori,A.,Mine,M.,Eshita,Y.: "Effects of high temperature on the emergence and survival of adult Culex pipiens molestus and Culex pipiens quinquefasciatus in Japan"J.Amer.Mosq.Cont.Assoc.. 15・20. 153-156 (1999)

    • Related Report
      2000 Annual Research Report
  • [Publications] 江下優樹,松本顕: "伝播昆虫の制御:遺伝子工学を用いた病原体耐性蚊の開発"医学のあゆみ. 191・1. 98-103 (1999)

    • Related Report
      2000 Annual Research Report
  • [Publications] 江下優樹: "研究最前線"大分医科大学学報. 78号. 18-20 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Eshita,Y.: "Vector competence of Japanese mosquitoes against dengue viruses"SP World. No.29. 13-17 (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] Takaoka,H.,Bain,O.,Uni,S.,Korenaga,M.,Tada,K.,Ichikawa,H.,Otsuka,Y.,and Eshita,Y.: "Human infection with an Onchocerca species from wild boars in Oita, Japan"Parasite. Vol.7(印刷中). (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] Oda,T.,Uchida,K.,Mori,A.,Mine,M.,Eshita,Y.: "Effects of high temperature on the emergence and survival of adult Culex pipiens molestus and Culex pipiens quinquefasciatus in Japan"J.Amer.Mosq.Cont.Assoc.. 15・20. 153-156 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] 江下優樹、松本 顕: "伝播昆虫の制御:遺伝子工学を用いた病原体耐性蚊の開発"医学のあゆみ. 191・1. 98-103 (1999)

    • Related Report
      1999 Annual Research Report

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Published: 1999-04-01   Modified: 2016-04-21  

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