利用绿色荧光蛋白及其变体评估杆状病毒感染
Evaluating Baculovirus Infection Using Green Fluorescent Protein and Variants.
作者信息
Wu Hsuan-Chen, Cha Hyung Joon, Bentley William E
机构信息
Department of Biochemical Science and Technology, National Taiwan University, Taipei, 10617, Taiwan.
Institute for Bioscience and Biotechnology Research, University of Maryland, College Park, MD, USA.
出版信息
Methods Mol Biol. 2016;1350:447-59. doi: 10.1007/978-1-4939-3043-2_22.
By use of a strategy incorporating the green fluorescent protein (GFP), facile and rapid monitoring and visualization of baculovirus infection in insect cells is possible in vivo. This chapter describes two techniques for simple determination of virus titer in the baculovirus expression system using GFP co-expression and rapid monitoring of Sf-9 insect cell infection using a combination of GFP and the early-to-late (ETL) promoter of the virus vector. Because of its early appearance, GFP, when placed under the control of ETL promoter, will facilitate vector construction, virus isolation, and titer determination.
通过使用一种结合绿色荧光蛋白（GFP）的策略，可以在体内轻松、快速地监测和可视化昆虫细胞中的杆状病毒感染。本章介绍了两种技术，一种是利用GFP共表达在杆状病毒表达系统中简单测定病毒滴度，另一种是结合GFP和病毒载体的早期到晚期（ETL）启动子快速监测Sf-9昆虫细胞感染。由于GFP出现较早，当置于ETL启动子控制下时，将有助于载体构建、病毒分离和滴度测定。
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