1989 Fiscal Year Final Research Report Summary
An attempt to produce virus resistant mice by manipulating mouse embryos
| Project/Area Number |
63580041
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Laboratory animal science
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| Research Institution | National Institute of Neuroscience, NCNP |
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Principal Investigator |
HANAOKA K. National Institute of Neuroscience, NCNP. Chief Researcher, 神経研究所・モデル動物開発部, 室長 (40189577)
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| Co-Investigator(Kenkyū-buntansha) |
TAGUCHI F. National Institute of Neuroscience, NCNP. Chief Researcher, 神経研究所・モデル動物開発部, 室長 (30107429)
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| Project Period (FY) |
1988 – 1989
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| Keywords | MHV / E2 protein / Chimeric mice / ES cell / gene transfer / ウイルス抵抗性動物 / E_2蛋白 / 細胞障害性 |
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| Research Abstract |
The mice used for experiments are bred free from the bacterial and viral infections which deteriorate the results of the animal experiments. These animals catled specific pathogen free (SPF) are therefor very sensitive to the infections. It has been often documented that SPF mouse colonies are attacked by the infection with mouse hepatitis virus (MHV) or sendai virus, which spoils the result of animal experiments completely. To overcome such problems, we have undertaken research to make the SPF animals resistant to MHV by gene transfer. MHV is known to infect cell via binding to the receptor on cell surface and E2 protein on virion surface is believed to be involved in that event. Our strategy to produce resistant animals to MHV is to cover the receptors on the susceptible cell surface by expressing E2 proteins in cells. The obstacle of this experiment is that the E2 protein itself is cytotoxic. Therefore, the domain involved in such cytottoxic activity must be excluded from E2 gene, or alternatively, the domain involved in the binding to the receptor should be isolated. To localize such domains, we first of all have expressed whole E2 gene in mouse L cells, which resulted in the death of cells expressing E2 and we failed to obtain the cell line expressing E2 continuousiy. Therefore, we have carried out the expression of E2 protein in insect cells by the baculovirus expression (BV) vectors. The E2 protein expressed by the recombinant BV showed high similarity to E2 protein produced in MHV infected cells in terms of immunogenicity, however, the protein lacked the activity to bind the receptor on mouse cells. This might be due to the difference in glycosilation of the E2 protein produced in insect cells. We are now trying to express E2 protein by vaccinia virus vectors in mouse cells in order to obtain the E2 protein with receptor binding activity.
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