Using fluorescent proteins to study poxvirus morphogenesis.
Using fluorescent proteins to study poxvirus morphogenesis.
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使用荧光蛋白研究痘病毒形态发生。
DOI：
10.1007/978-1-59745-559-6_1
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发表时间：
2009
期刊：
影响因子：
--
通讯作者：
Ward,BrianM
中科院分区：
文献类型：
--
作者：
Ward,BrianM
Fluorescent protein (FP) fusions not only allow for the convenient visualization of a protein of interest’s subcellular localization but also permit the real-time monitoring of their subcellular trafficking. The subcellular fluorescent pattern of FP-fusions can also serve as a visual marker for various subcellular processes using either live or static microscopy. We have employed FP-fusions for the study of poxvirus morphogenesis. Fusion of FP with either a viral core protein or an extracellular virion-specific protein can serve as a visual read-out for normal poxvirus morphogenesis at the subcellular level. Recombinant viruses expressing a FP-fusion, in conjunction with the deletion of a gene involved in either morphogenesis or egress, usually display an aberrant FP pattern. Functional domains in the missing protein are then mapped by complementation in-trans followed by fluorescent microscopy for analysis of the FP pattern. The methods presented here describe how to infect and transfect cells for trans-complementation for the purpose of functional domain mapping. The imaging and analysis of these cells is described.
