| Project/Area Number |
18590445
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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 |
Virology
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| Research Institution | Hokkaido University |
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Principal Investigator |
KANDA Teru Hokkaido University, Inst. for Genetic Med., Assistant Professor (50333472)
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| Project Period (FY) |
2006 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
|
| Budget Amount *help |
¥4,040,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥540,000)
Fiscal Year 2007: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
Fiscal Year 2006: ¥1,700,000 (Direct Cost: ¥1,700,000)
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| Keywords | Epstein-Barr virus / bacterial artificial chromosome / packaging system / 293 cells / B-lymphocyte transformation |
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| Research Abstract |
Bacterial artificial chromosome (BAC) system is useful for engineering the genome of Epstein-Barr virus (EBV) genome in E. coll. We aimed to establish a packaging system to produce recombinant EBVs derived from the BAC clone. We tested two cell lines harboring helper viruses of EBV for their usefulness as packaging cells; (1)P3HR-1 cells (derived from human Burkitt's lymphoma cells) and (2)B95-8 cells and their derivatives (derived from marmoset lymphoblastoid cell lines). However, the results revealed that we could not avoid the recombination between the helper virus DNA and the BAC clone DNA. Therefore, in order to get pure recombinant EBVs, we switched our strategy to establish a system that is free from helper virus. For this purpose, we cloned the full length genome of B95-8 strain EBV, and introduced the BAC clone DNA into human 293 cells. This strategy enabled us to efficiently establish recombinant virus-producing cells. The resultant recombinant EBVs are superior to previously-reported 293-derived recombinant EBVs in their efficiency to transform B-lymphocyte as well as in their ability to express transgenes. These results demonstrate that the combinational usage of the BAC clone of B95-8 strain EBV and 293 cells is an efficient way to produce high-titer pure recombinant EBVs.
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