Establishment of a GFP-based indicator cell line to quantitate feline foamy virus

被引:10
|
作者
Phung, HTT
Tohya, Y
Shimojima, M
Kato, K
Miyazawa, T
Akashi, H
机构
[1] Univ Tokyo, Grad Sch Agr & Life Sci, Dept Vet Microbiol, Bunkyo Ku, Tokyo 1138657, Japan
[2] Osaka Univ, Microbial Dis Res Inst, Res Ctr Emerging Infect Dis, Suita, Osaka 5650871, Japan
[3] Japan Sci & Technol Corp, PRESTO, Tokyo, Japan
关键词
feline foamy virus; titration; GFP; transactivation; cell-tropism;
D O I
10.1016/S0166-0934(03)00062-4
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
To quantitate infectious feline foamy virus (FeFV), Crandell feline kidney (CRFK) cells were transfected with the gfp gene under the control of the FeFV long terminal repeat (LTR) for establishing an indicator cell line named FFG cells. The FeFV activates promoter activity of the LTR to express green fluorescent protein (GFP) upon infection. The titers determined by GFP-positive FFG cells (GFP-based assay) were higher than those determined by the cytopathic effects-positive CRFK cells (CPE-based assay). The titers determined by the GFP-based assay reached a plateau at 3-4 days post infection (d.p.i.), while those by the CPE-based assay reached 6-8 d.p.i. When stock viruses of various FeFV strains were titrated by both assays, titers determined by both assays correlated well with each other. The results show that the GFP-based assay is simpler and more rapid and sensitive than the CPE-based assay. Using the GFP-based assay, we examined the in vitro host range of FeFV. It was found that FeFV can productively infect various cell lines derived from cats, dogs, chickens, a human and a bat. (C) 2003 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:125 / 131
页数:7
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