| Project/Area Number |
10670508
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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 |
Gastroenterology
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| Research Institution | Tokai University |
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Principal Investigator |
KAGAWA Tatehiro Tokai University, School of Medicine, Assistant Professor, 医学部, 講師 (30245469)
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| Co-Investigator(Kenkyū-buntansha) |
HISOI Katsumi Tokai University, School of Medicine, Assistant Professor, 医学部, 助手 (50317814)
NISHIZAKI Yasuhiro Tokai University, School of Medicine, Assistant Professor, 医学部, 講師 (80237693)
WATANABE Norihito Tokai University, School of Medicine, Associate Professor, 医学部, 助教授 (90167156)
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| Project Period (FY) |
1998 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 1999: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1998: ¥2,400,000 (Direct Cost: ¥2,400,000)
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| Keywords | p-glycoprotein / hepatocyte / multidrug resistance / green fluorescent protein / intracellular trafficking / P-glycoprotein / sorting / 肝細胞 |
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| Research Abstract |
1. Construct of expression vector
We made two types of vectors which expressed a fusion protein of human p-glycoprotein (human multidrug-resistance 1, hMDR1) and green fluorescent protein (GFP). One of them was made to express GFP-hMDR1, the other hMDR1-GFP.
2. Western blotting
These vectors were transfected into HeLa cells and the cell lysate was served for Western blotting. We could detect a reaction of 220 kD using anti-GFP and C219 (anti-p-glycoprotein), suggesting that these vectors could intracellularly express fusion proteins of GFP and hMDR1.
3. Localization of fusion protein
The localization of the GFP-hMDR1 fusion protein was diffuse in various types of cells, which result was the same as GFP alone. On the other hand, hMDR1-GFP was found along the plasma membrane. Therefore, this vector, which expresses the hMDR1-GFP protein, would be very useful to investigate intracellular trafficking mechanism.
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