| Project/Area Number |
11557188
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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 | 展開研究 |
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| Research Field |
医薬分子機能学
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| Research Institution | Tohoku University |
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Principal Investigator |
ABE Takaaki Tohoku Univ., School of Med., Neurophysiology, Research Associate, 大学院・医学系研究科, 助手 (80292209)
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| Co-Investigator(Kenkyū-buntansha) |
YAWO Hiromu Tohoku Univ., School of Med., Neurophysiology, Porfessor, 大学院・医学系研究科, 教授 (00144353)
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| Project Period (FY) |
1999 – 2000
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| Project Status |
Completed (Fiscal Year 2000)
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| Budget Amount *help |
¥13,500,000 (Direct Cost: ¥13,500,000)
Fiscal Year 2000: ¥6,700,000 (Direct Cost: ¥6,700,000)
Fiscal Year 1999: ¥6,800,000 (Direct Cost: ¥6,800,000)
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| Keywords | organic anion transporter / Thyroid hormone transporter / adenovirus / blood-brain-barrier / choroid plexus / gene transfer / トランスポーター / 有機アニオン / プロスタグランジン / 胆汁酸 / 海馬 |
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| Research Abstract |
We have established adenovirus-mediated gene transfer system into cells.
The coding region of LST-1 and LST-2, both of which have isolated by us have introduced into adenovirus vector, packaged, and transfected into HEK293 cell with adenovirus genomic DNA.The cells that produces homologous-recombinant shrieked and made plaque formation. After producing large scale infected medium, the recombinant adenovirus was purified by CsCl ultracentrifuge.
The infection titer of the purified LST-1 and LST-2 adenoviruses were measured and the IC 50 of each virus was 1.26 and 1.48 X 10^<10> pfu, respectively.
When recombinant virus was applied to cultured cell line, the LST-1 and LST-2 protein was strongly expressed within 24 hr after application. The size of the band was also confirmed by Western blot analysis. The LST-1 and LST-2 expressing mammalian cell transports various organic anions as substrates and the apparent Km value of tauro-cholate uptake is 24 micro M and 32 micro M, respectively and both of which is a comparative value of the native cells reported.
The establishment of the adenovirus-mediated gene transfer system wiil lead to the various applied sysem for screening of the brain-, liver and kidney oriented drug delivery and therapeutic tools.
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