| Project/Area Number |
15109008
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| Research Category |
Grant-in-Aid for Scientific Research (S)
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| Allocation Type | Single-year Grants |
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| Research Field |
Digestive surgery
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| Research Institution | The University of Tokyo |
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Principal Investigator |
MAKUUCHI Masatoshi The University of Tokyo, Faculty of Medicine, Professor, 医学部附属病院, 教授 (60114641)
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| Co-Investigator(Kenkyū-buntansha) |
NARUSE Katsutoshi The University of Tokyo, Faculty of Medicine, Assistant, 医学部附属病院, 助手 (50291323)
NAGASHIMA Hiroshi The Meiji University, Faculty of Agriculture, Professor, 農学部, 教授 (50318664)
東條 英昭 東京大学, 大学院・農学生命科学研究科, 教授 (20041668)
澤崎 徹 東京大学, 大学院・農学生命科学研究科附属牧場, 教授 (00012047)
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| Project Period (FY) |
2003 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥118,820,000 (Direct Cost: ¥91,400,000、Indirect Cost: ¥27,420,000)
Fiscal Year 2005: ¥38,740,000 (Direct Cost: ¥29,800,000、Indirect Cost: ¥8,940,000)
Fiscal Year 2004: ¥38,350,000 (Direct Cost: ¥29,500,000、Indirect Cost: ¥8,850,000)
Fiscal Year 2003: ¥41,730,000 (Direct Cost: ¥32,100,000、Indirect Cost: ¥9,630,000)
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| Keywords | Transgenic pig / Albumin / EGFP / ヒト血清アルブミン / 人工肝臓 / 人血清アルブミン / 動物バイオリアクター |
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| Research Abstract |
Producing transgenic cattle that can secret human proteins is a promising strategy for development of a bioreactor for human pharmaceuticals or bioartificial liver support. We constructed pCX-hAlb-EGFP, that is, a combined gene of human albumin cDNA and EGFP cDNA connected with CMV-IE enhancer, chicken β-actin promoter, and rabbit β-globin poly A signal for the purpose of producing a transgenic pig that can secret human albumin systemically. NIH3T3 cells transformed with pCX-hAlb-EGFP by the lipofection method showed intense expression of human albumin and EGFP genes. Thus, we introduced it into porcine oocytes using sperm vector method, and a piglet was produced which showed clear expression of GFP in the hooves and skin. PCR analysis of genomic DNA extracted from several tissues including the liver showed that they possessed the transgene. RT-PCR analysis demonstrated that both the human albumin and EGFP genes were expressed in the same tissues. The transgene was integrated and expressed in the porcine liver, which is indispensable for our purpose of utilizing the transgenic liver for bioartificial liver support. Real time RT-PCR analysis exhibited that the expression rate of human albumin gene and EGFP gene against β-actin promoter in the liver showed adequate quantity of transcribed mRNA and fairly good expression rates as compared with those of the other tissues. These results will promise successful result of further trials of producing desirable transgenic pig with the liver secreting human albumin.
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