Expression of erythrocyte binding protein in Plasmodium falciparum merozoites isolated from endemic area
| Project/Area Number |
13576007
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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 | 海外学術 |
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| Research Field |
寄生虫学(含医用動物学)
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| Research Institution | Ehime University |
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Principal Investigator |
TORII Motomi Ehime University, Faculty of Medicine, Professor, 医学部, 教授 (20164072)
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| Co-Investigator(Kenkyū-buntansha) |
MATSUDA Syouji Ehime University, Faculty of Medicine, Professor, 医学部, 教授 (40173843)
KANEKO Osamu Ehime University, Faculty of Medicine, Instructor, 医学部, 助手 (50325370)
TSUBOI Takafumi Ehime University, Faculty of Medicine, Associate Professor, 医学部, 助教授 (00188616)
IRIKO Hideyuki Ehime University, Faculty of Medicine, Research Assistant, 医学部, 教務職員 (60346674)
橘 真由美 愛媛大学, 医学部, 教務職員 (00301325)
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| Project Period (FY) |
2001 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥13,600,000 (Direct Cost: ¥13,600,000)
Fiscal Year 2002: ¥4,500,000 (Direct Cost: ¥4,500,000)
Fiscal Year 2001: ¥9,100,000 (Direct Cost: ¥9,100,000)
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| Keywords | infectious disease / malaria / Plasmodium / invasion / protein expression / RT-PCR / parasitology / 赤血球 |
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| Research Abstract |
Mutant erythrocyte that lacked certain proteins on the surface were frequently observed in the malaria endemic areas, and some of them were believed to be generated by the selective advantage against malaria infection. However, Plasmodium falciparum have an ability to invade many types of erythrocyte including these mutant erythrocytes. The molecular base of this ability of P.falciparum could be resulted by the erythrocyte-binding proteins encoded in the multi-gene family involved in the invasion steps. In this study, we explored the possibility if P.falciparum regulated the expression of these multi-gene family in order to overcome the erythrocyte polymorphism by quantitate the transcription and protein expression level of each members of the multi-gene family, PfRhopH1.
Firstly, we designed a panel of oligonucleotide primers for real-time PCR method with SYBR green. Complementary DNA was made from the 3D7 clone of P.falciparum and used to make plasmids containing the real-time PCR targets for each member. These plasmids were used to create the standard curve. P.falciparum field samples were collected in the malaria endemic area in Thailand and total RNA were extracted. Simultaneously blood smear were made on the glass slides for the IFA analysis for the protein expression. In order to check the protein expression level, we successfully generated anti-PfRhopHl_2, 3.1, 9 specific sera. Difference in the protein expression was observed among the culture-adapted P. falciparum clones.
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Report
(3 results)
Research Products
(7 results)
