| Project/Area Number |
09670268
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
寄生虫学(含医用動物学)
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| Research Institution | Kumamoto University |
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Principal Investigator |
MIMORI Tatsuyuki Kumamoto University, School of Medicine, Associate Professor, 医学部, 助教授 (00117384)
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| Project Period (FY) |
1997 – 1998
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| Project Status |
Completed (Fiscal Year 1998)
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| Budget Amount *help |
¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 1998: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1997: ¥2,300,000 (Direct Cost: ¥2,300,000)
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| Keywords | Leishmania parasite / Leishmania amazonensis / Molecular biotechnology / Invasion / Genomic DNA library / Deletion mutant / Non-pathogenic E.coli / Kinetoplast DNA / 細胞侵入 / kinetoplast(k)DNA / DNAライブラリー |
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| Research Abstract |
Leishmaniases are widely distributed in tropical and temperate zones, where approximately 12 million people are affected. However, there is not drugs for appropriate treatment of Leishmania parasites. Leishmania parasitizes to the mammalina cells. Invasion mechnisum of this parasite are not known yet. We searched the Leishmania invasion molecule using genome and cDNA library and non-pathogenic E.coil system. Genome and cDNA were made cultured promasigotes of Leishmania amazonensis. These DNAs were partially digested with Eco RI and Sau 3A1, then were ligated to plasmid vector (pBluescript IIsk+). These plasmid cotaining Leishmania DNA insert were infected into E.coli (DH5alpha). The E.coli were incubated with mammalian cells (VA-13). Invaded bacteria were selected with washing of antibiotic fluid. Five clones of selected 11 clones had the same 614 base pairs sequences. Furthermore, we made deletion mutants of this insert Leishmania DNA.From this experiment, it was suggested that about 130 base pairs sequences N-terminal was more important site for invasion to the host cell. The GST-fusion protein was made using plasmid (pGEX-2TH) and E.coli (NM522) system. Adhesion activity of this protein was about 20 times compared with control one.
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