| Project/Area Number |
04044111
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| Research Category |
Grant-in-Aid for international Scientific Research
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| Allocation Type | Single-year Grants |
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| Section | Joint Research |
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| Research Institution | Research Institute for Microbial Diseases, Osaka University |
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Principal Investigator |
SHIMADA Kazunori Res.Inst.for Microbial Diseases, Osaka University, 微生物病研究所, 教授 (40037354)
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| Co-Investigator(Kenkyū-buntansha) |
COSTANTINI F コロンビア大学, 医学部, 教授
GOTTESMAN Ma コロンビア大学, 癌研究所, 所長
MAEDA Syuichiro Yamanashi Medical University, 医学部, 教授 (10117244)
NISHIGUCHI Seiji Res.Inst.for Microbial Diseases, Osaka University, 微生物病研究所, 助手 (90237686)
JOH Tadashi Res.Inst.for Microbial Diseases, Osaka University, 微生物病研究所, 助手 (50243259)
TAKIHARA Yoshihiro Res.Inst.for Microbial Diseases, Osaka University, 微生物病研究所, 助教授 (60226967)
COSTANTINI Franklin d. Department of Genetics and Development, Columbia University
CONSTANTINI エフ.ディー. コロンビア大学, 医学部, 教授
ROBERTSON E. ハーバード大学, 教授
F D Constant コロンビア大学, 医学部, 教授
E J Robertso ハーバード大学, 教授
MAX E Gottes コロンビア大学癌研究所, 所長
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| Project Period (FY) |
1992 – 1994
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| Project Status |
Completed (Fiscal Year 1994)
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| Budget Amount *help |
¥14,700,000 (Direct Cost: ¥14,700,000)
Fiscal Year 1994: ¥4,900,000 (Direct Cost: ¥4,900,000)
Fiscal Year 1993: ¥4,900,000 (Direct Cost: ¥4,900,000)
Fiscal Year 1992: ¥4,900,000 (Direct Cost: ¥4,900,000)
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| Keywords | ES / FAP / Transthyretin / TTR-deficient mouse / Retinol / RBP / Thyroid hormone / Replacement type vector / 血清アミロイドP成分 |
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| Research Abstract |
1.To generate a new mouse model of familial amyloidotic polyneuropathy (FAP) and to elucidate the function of the human variant TTR,we introduced the human mutant transthyretin (ttr) gene into the TTR-deficient mice. In the TTR-deficient mice carrying the human mutant ttr gene, amyloid began to deposit at the age 11 months. We have not yet observed any significant differences in the onset, progression and tissue distribution of amyloid deposition between the wild type and TTR-deficient transgenic mice carrying the human mutant gene. These mice have not developed peripheral neuropathy yet.
2.The introduction of the human mutant gene into the TTR-deficient mice increased the serum level of retinol-binding protein (RBP) from 3% to 84% of the wild type mice. The introduction also increased the serum level of thyroid hormone (T4) from 40% to 56% of the wild type mice.
3.To produce mice carrying a point mutation in the ttr gene, we developed a novel replacement type vector. This vector consists of a part of the ttr gene carrying the point mutation in its 2nd exon, and a cassette of selection markers (MC1-neo and MC1-tk) flanked with a 3 kb duplication of the 2nd intron of the ttr gene. After electroporating the vector into ES cells, 2 targeted clones carrying the selection markers as well as the point mutation in the ttr gene were isolated from 300 G418-resistant clones. Sixteen FIAU-resistant clones were isolated from one of the two targeted clones. Two of the 16 clones carried no selection markers, but retained the point mutation in the ttr gene. After injecting the targeted ES cells into C57BL/6 host blastocysts, chimeric mice were obtained. The chimeric mice were now being bred with C57BL/6 females. These results suggested that this vector is useful to introduce subtle mutations efficiently into mouse genes.
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