1986 Fiscal Year Final Research Report Summary
Expression of mouse IL2 gene and its mass production by using virus vector-mammalian cell line
| Project/Area Number |
60570216
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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 |
Immunology
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| Research Institution | Chiba University |
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Principal Investigator |
FUSE Akira Chiba University, School of Medicine,Research Associate, 医学部, 助手 (60110300)
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| Project Period (FY) |
1985 – 1986
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| Keywords | Mouse IL2 / Gene Expression / Virus Vector / パピローマウィルス |
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| Research Abstract |
To characterize the biological and chemical properties of mouse interleukin 2 (IL2) I have tried to clone mouse IL2 cDNA and express IL2 by using the bovine papillomavirus vector and mouse cultured cell line. Mouse IL2 cDNA was cloned from the cDNA library made from activated spleen cells by concanavalin A. The IL2 cDNA was inserted downstream of SV40 late promotor in bovinepapillomavirus vector (BPV1). The constructed IL2-BPV DNA was transfected to C127 cell by Ca-phosphate precipitate method. Three weeks after transfection 7 transformant cell lines were cloned. The amounts of IL2 produced in culture medium were 3-15 units/ml. These cell lines continued to produce IL2 for several months, but reduced its production after 6 months. Two of 7 cell lines stopped the production of IL2. The copy numbers of each cell lines were 35-200/cell just after cloning and 0-70/cell after 6 months, respectively. Two cell lines showed the integration of IL2-BPV DNA into host genome produced very low amounts of IL2. It is suggested to develop more stable BPV vector and introduce stronger promotor for the long term production and higher production, respectively.
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