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Mechanisms of drug-resistance and analysis of drug-resistant genes in Leishmania

Research Project

Project/Area Number 05670243
Research Category

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

Allocation TypeSingle-year Grants
Research Field 寄生虫学(含医用動物学)
Research InstitutionJikei University School of Medicine

Principal Investigator

KATAKURA Ken  Jikei University School of Medicine, Department of Tropical Medicine, Instructor, 医学部, 講師 (10130155)

Project Period (FY) 1993 – 1994
Project Status Completed (Fiscal Year 1994)
Budget Amount *help
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1994: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1993: ¥1,600,000 (Direct Cost: ¥1,600,000)
KeywordsLeishmania / drug resistance / P-glycoproteins / polymerase chain reaction / multigene family / DNA / parasitic protozoa / pulsed field gel electrophoresis / P‐糖タンパク質 / 分子生物学 / 寄生性原虫
Research Abstract

A protozoan parasite, Leishmania, contains several P-glycoprotein genes as a multigene family. Two types of the gene (Ldmdrl and LtpgpA) have been isolated from different Leishmania species, and found to play a role of drug-resistance. In this study, we aimed to isolate and characterize P-glycoprotein genes in L.amazonensis, one of the causative agents of cutaneous leishmaniasis in the New World. Polymerase chain reaction (PCR) was performed to amplify ATP binding sites within P-glycoproteins of L.amazonensis by using primers specific to conserved sequences in mammalian and leishmanial P-glycoprotein genes. We detected a 408-bp PCR product which showed 68% and 37% identity to Ldmdrl and LtpgpA,respectively, at the amino acid level, suggesting that this DNA fragment is in part of new type of leishmanial P-glycoprotein gene. To isolate the putative gene in whole, a genomic library of L.amazonensis DNA was constructed into the lambda-GEM-11 vector and screened with the PCR product. One of the positive lambda clones was digested with Sac I restriction endonuclease, and resultant DNA fragments were subcloned into pUC19 vector. At present, nucleotide sequencing of a 1005-bp Sac I fragment was completed, and the rest of the fragments is ready to be sequenced. In addition, pulsed field gel electrophoresis and Southern hybridization analysis strongly suggest that the L.amazonensis contains at least three different types of P-glycoprotein gene, the Ldmdrl homologue, LtpgpA homologue and new type gene found in this study, but that chromosomal location of the LtpgpA is different from those of the other two genes.

Report

(3 results)
  • 1994 Annual Research Report   Final Research Report Summary
  • 1993 Annual Research Report
  • Research Products

    (19 results)

All Other

All Publications (19 results)

  • [Publications] Katakura,K.: "Karyotype similarity of Leishmania isolates from patients,sandflies, anda domestic dog,identifying the major L. (L.)mexicana strain as an agent of cutaneous leishmaniasis in the Ecuadorian Andes." Studies on New World leishmaniasis and its trans- mission,with particular reference to Ecuador.4. 4-8 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Katakura, K.: "Evaluation of Glucantime (Meglumine Antimonate)lots in anti-Leishmania promastigote activity in vitro." Studies on New World leishmaniasis and its trans-mission, with particular reference to Ecuador.4. 30-33 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 大友弘士: "免疫検査実践マニュアルVIII感染症5.原虫疾患" 検査と技術. 22. 312-314 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 大友弘士: "微生物のVIRULENCE原虫" Clinical Infection and Chemotherapy. 1. 38-41 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 片倉賢: "リーシュマニアの薬物療法とリーシュマニアの薬剤耐性" メディアサークル. (印刷中). (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Katakura, K.: "Karyotype similarity of Leishmania isolates from patients, sandflies, and a domestic dog, identifying the major L.(L.) mexicana strain as an agent of cutaneous leishmaniasis in the Ecuadorian Andes. In Studies on New World leishmaniasis and its transmission, with particular reference to Ecuador. Research Report Series No.4. Hashiguchi, Y., ed. Kyowa Printing, Kochi." 4-8 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Katakura, K.: "Evaluation of Glucantime(]SY.encircledR.[) (Meglumine Antimonate) lots in anti-Leishmania promastigote activity in vitro. In Studies on New World leishmaniasis and its transmission, with particular reference to Ecuador. Research Report Series No.4. Hashiguchi, Y., ed. Kyowa Printing, Kochi." 30-33 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Ohtomo, H.: "Practical manuals for immunological tests. VIII.Infectious diseases. 5. Parasitic diseases by protozoa infection (in Japanese)." Modern Medical Laboratory. 22. 312-314 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Ohtomo, H.: "Virulence of microorganisms : parasitic protozoa. (in Japanese)" Clinical Infection and Chemotherapy. 1. 38-41 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Katakura, K.: "Chemotherapy of leishmaniasis and drug-resistance in Leishmania (in Japanese)." Media-Circle. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Katakura,K.: "Karyotype similarity of Leishmania isolates from patients,sandflies,and a domestic dog,identifying the major L. (L.) mexicana strain as an agent of cutaneous leishmaniasis in the Ecuadorian Andes." Studies on New World leishmaniasis and its transmission,with particular reference to Ecuador. 4. 4-8 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Katakura,K.: "Evaluation of Glucantime (Meglumine Antimonate) lots in anti-Leishmania promastigote activity in vitro." Studies on New World leishmaniasis and its transmission,with particular reference to Ecuador.4. 30-33 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] 大友弘士: "免疫検査実践マニュアルVIII感染症5.原虫疾患" 検査と技術. 22. 312-314 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] 大友弘士: "微生物のVIRULENCE原虫" Clinical Infection and Chemotherapy. 1. 38-41 (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] 片倉賢: "リーシュマニア症の薬物療法とリーシュマニアの薬剤耐性" メディアサークル. (印刷中). (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] Katakura,K.et al.: "Molecular karyotype characterization of Leishmania panamensis,L.mexicana and L.major-like parasites:agents of cutaneous leishmaniasis in Ecuador." American Journal of Tropical Medicine and Hygiene. 48. 707-715 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Katakura,K.et al.: "Cutaneous leishmaniasis in mast cell-deficient W/W^V mice." Infection and Immunity. 61. 2242-2244 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] 片倉 賢: "リーシュマニア症の伝播疫学とリーシュマニア原虫の生態" 日本熱帯生態学会ニューズレター. 12. 6-9 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] 元木良和: "Cutaneous leishmaniasis" 臨床皮膚科. 5. 17-21 (1993)

    • Related Report
      1993 Annual Research Report

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Published: 1993-04-01   Modified: 2016-04-21  

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