| Project/Area Number |
12670289
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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 |
Virology
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| Research Institution | The University of Tokyo (2001-2002)
National Institute of Infectious Diseases (2000) |
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Principal Investigator |
KITAMURA Yoshihiro The University of Tokyo, INSTITUTE OF MEDICAL SCIENCE, Associate Professor, 医科学研究所, 助教授 (10202037)
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| Project Period (FY) |
2000 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 2002: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2001: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2000: ¥1,500,000 (Direct Cost: ¥1,500,000)
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| Keywords | human immunodeficiency virus / INTEGRATION / INTEGRACE / VECTOR / ヒト免疫全ウルス / HIV / ini1 / SNF5 / アポトーシス / G1アレスト / 組み込み酵素 / 単クローン抗体 |
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| Research Abstract |
To examine whether vectors based on human immunodeficiency virus type 1 (HIV) with viral central polypurine tract (cPPT) express transgenes more efficiently than those without cPPT, we constructed a series of vectors carrying a Neo'gene as a stable marker, which is driven by an internal synthetic thymidine kinase promoter. Insertion of cPPT, either 282bp or 178bp long, resulted in about 3-8-fold increase of the amount of produced vector particles. Insertion of the longer cPPT. resulted in decrease in transduction efficiency in about eight-fold, whereas insertion of the shorter resulted in increase in transduction efficiency in about two-fold independently of the orientation of the insert. Moreover, DNA-PCR showed that reverse transcription and nuclear import were cnot affected. Alu-PCR assay showed that integration was impaired in the vectors carrying the 282bp-cPPT. Taking these together, we concluded that insertion of the 282bp-cPPT decreased the number of 0418-resistant colonies without affecting nuclear import of vector DNA. We suppose that insertion of the 282bp-cPPT resulted in impairment of events after import, such as integration.
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