| Project/Area Number |
08456149
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Basic veterinary science/Basic zootechnical science
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| Research Institution | The University of Tokyo |
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Principal Investigator |
MATSUMOTO Yoshitsugu Univ.of Tokyo, Grad.Sch.of Agri.and Life Sci., Associate Professor, 大学院・農学生命科学研究科, 助教授 (00173922)
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| Co-Investigator(Kenkyū-buntansha) |
KAWAZU Shin-ichiro Natl.Inst.Anim Health, Researcher, 家畜衛生試験場, 主任研究員
ONODERA Takashi Univ.of Tokyo, Grad.Sch.of Agri.and Life Sci., Professor, 大学院・農学生命科学研究科, 教授 (90012781)
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| Project Period (FY) |
1996 – 1997
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| Project Status |
Completed (Fiscal Year 1997)
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| Budget Amount *help |
¥7,400,000 (Direct Cost: ¥7,400,000)
Fiscal Year 1997: ¥2,900,000 (Direct Cost: ¥2,900,000)
Fiscal Year 1996: ¥4,500,000 (Direct Cost: ¥4,500,000)
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| Keywords | IL-8 / cDNA / expression vector / Leishmania / parasite / canine / 発現用ベクター / cDNA |
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| Research Abstract |
Leishmaniasis is often a zoonosis in which dog is known to be one of the most important animal reservoirs. For control of such zoonotic deseases, development of a vaccine directed against the natural reservoir host is desirable. In this study, we successfully produced, by molecular engineering, Leishmania amazonensis promastigotes expressing biologically active canine interleukin-8 (cIL-8). The PCR products of cIL-8 cDNA were inserted into the L.amazonensis expression vector (p6.5) which was designed to contain a tunicanycin (TM) resistance gene for selection. The constructed expression vector (p6.5-cIL-8) was transfected into promastigotes of a L.amazonensis clone (C_1/12) by electroporation. Transfectants were selected in culture madium with TM.By PCR of extrachromosomal DNAs from the transfectants using cIL-8 specific primers, a predicted product of a 330 bp long was amplified. Western blot analysis of the trnsfectants demonstrated a peptide of about 8.8kDa in molecular size, as expected of cIL-8. The culture supernatant of the transfectant showed a chemotactic activity with dog and mouse polymorphonuclear cells (PMN).
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