| Project/Area Number |
07670601
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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 |
Gastroenterology
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| Research Institution | Kumamoto University |
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Principal Investigator |
FUJIYAMA Shigetoshi Kumamoto University, School of Medicine, Associate Professor, 医学部, 助教授 (20109656)
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| Co-Investigator(Kenkyū-buntansha) |
CHIKAZAWA H Kumamoto University, Hospital, Clinical Fellow, 医学部・附属病院, 医員
SUGI K Kumamoto University, , School of Medicine Assistant Professor, 医学部, 助手 (00274704)
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| Project Period (FY) |
1995 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1996: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1995: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Keywords | hepatitis C / HCV / gene therapy / antisense |
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| Research Abstract |
1) Construction in vitro transcription system based on recombinant RNA polymerase derived HCV genome for establishment of native HCV-RNA trnscription system.
AcDNA corresponding to the NS5-coding region has been isolated and cloned, which region is coding HCV own RNA polymerase. Construction of translation system of recombinant RNA polymerase derived HCV genome has been tried by using E.coli or yeast, but failed. Continuously, construction in vitro HCV RNA transcription system based on both T7 promoter and T7 RNA polymerase has been tried, finally HCV RNA failed to be transcribed.
2) A study of suppression of HCV translation by using antisense genome.
Expression of HCV recombinant protein considered as core-envelope protein was achieved, based on recombinant plasmid, including HCV genome corresponding to 5'-noncoding region, core region and part of envelope region. In this expression system, antisense oligonucleotide targeting near by starting codon mapped on 5'-noncoding region was added to study inhibitory effect of antisense genome to translation of HCV genome. Consequently, the highly inhibitory effect, which showed dose depending, has confirmed. Nevertheless, some inhibitory effect of sense genome to translation was observed. It was accordingly suggested that the antisense oligonucleotide non-specifically suppressed HCV translation. We are trying to find the cause of non-specific effect of antisense genome and the best plan. Some revision are needed in the method of in vitro translation and modification of antisense oligonucleotide.
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