| Project/Area Number |
12670288
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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 | NATIONAL INSTITUTE OF INFECTIONS DISEASES |
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Principal Investigator |
TAKEBE YUTAKA NATIONAL INSTITUTE OF INFECTIONS DISEASES, AIDS RESEARCH CENTER, CHIEF, エイズ研究センター第1室, 室長 (50126116)
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| Project Period (FY) |
2000 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2001: ¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 2000: ¥1,800,000 (Direct Cost: ¥1,800,000)
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| Keywords | HIV-1 subtype / HIV-1 envelope / Sendai virus vector / EIA / serotyping / HIV-1エンベロープタンパク質 / EIA / ELISA |
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| Research Abstract |
We developed Sendai virus (SeV) vectors that express HIV-1 env gp120s of subtypes A through E. In order to assess the expression level ofgp120, we developed the sandwich enzyme immunoassay (ElA) using purified polyclonal sera against C-terminus of gp120 (first antibody) and pooled HIV(+) sera (second antibody). Employing this assay system, we evaluated the effect of codon optimization and of fusion with CD5 leader sequence on expression level of gp120.We found that codon-optimization enhanced the expression of gp120 by 3-fold and the fusion to CD5 leader sequence showed additional enhancement of 1.5-fold.
EIA using the recombinant gp120 expressed by SeV vectors could differentiate the subtypes between B and E. Since high-level of cross-reactivity were observed between other subtypes, we found that the universal serological subtyping of HIV-1 subtypes is not feasible to achieve.
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