用于病毒-细胞相互作用分析的绿色荧光蛋白标记逆转录病毒包膜蛋白
Green fluorescent protein-tagged retroviral envelope protein for analysis of virus-cell interactions.
作者信息
Spitzer Dirk, Dittmar Kurt E J, Rohde Manfred, Hauser Hansjörg, Wirth Dagmar
机构信息
Department of Gene Regulation and Differentiation, German Research Center for Biotechnology, D-38124 Braunschweig, Germany.
出版信息
J Virol. 2003 May;77(10):6070-5. doi: 10.1128/jvi.77.10.6070-6075.2003.
Abstract
Fluorescent retroviral envelope (Env) proteins were developed for direct visualization of viral particles. By fusing the enhanced green fluorescent protein (eGFP) to the N terminus of the amphotropic 4070A envelope protein, extracellular presentation of eGFP was achieved. Viruses incorporated the modified Env protein and efficiently infected cells. We used the GFP-tagged viruses for staining retrovirus receptor-positive cells, thereby circumventing indirect labeling techniques. By generating cells which conditionally expressed the GFP-tagged Env protein, we could confirm an inverse correlation between retroviral Env expression and infectivity (superinfection). eGFP-tagged virus particles are suitable for monitoring the dynamics of virus-cell interactions.
摘要
荧光逆转录病毒包膜（Env）蛋白被开发用于病毒颗粒的直接可视化。通过将增强型绿色荧光蛋白（eGFP）融合到双嗜性4070A包膜蛋白的N端，实现了eGFP的细胞外展示。病毒整合了修饰后的Env蛋白并有效感染细胞。我们使用绿色荧光蛋白标记的病毒对逆转录病毒受体阳性细胞进行染色，从而避免了间接标记技术。通过生成条件性表达绿色荧光蛋白标记的Env蛋白的细胞，我们可以证实逆转录病毒Env表达与感染性（超感染）之间呈负相关。绿色荧光蛋白标记的病毒颗粒适用于监测病毒与细胞相互作用的动态过程。
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