A rapid method for titrating baculovirus stocks using the Sf-9 Easy Titer cell line
A rapid method for titrating baculovirus stocks using the Sf-9 Easy Titer cell line
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DOI：
10.2144/000113238
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发表时间：
2009-09-01
期刊：
影响因子：
2.7
通讯作者：
Esposito, Dominic
中科院分区：
文献类型：
--
作者：
Hopkins, Ralph F.;Esposito, Dominic
使用新型细胞系 Sf-9 Easy Titer (Sf-9ET) 开发了一种新的快速滴定杆状病毒储液的方法。 Sf-9ET 细胞系已用含有增强型绿色荧光蛋白 (eGFP) 基因的质粒 DNA 转染，该基因受杆状病毒多角体启动子控制。当用于滴定测定时，Sf-9ET细胞在被杆状病毒感染时变绿，这是由于感染期间表达的杆状病毒基因产物激活了多角体启动子/eGFP复合物。使用 96 孔板形式终点稀释测定，可以使用荧光显微镜在三天内测定杆状病毒滴度。
A new rapid method for titrating baculovirus stocks has been developed using a novel cell line Sf-9 Easy Titer (Sf-9ET). The Sf-9ET cell line has been transfected with plasmid DNA containing the enhanced green fluorescent protein (eGFP) gene under the control of the baculovirus polyhedrin promoter. When used in the titration assay, the Sf-9ET cells turn green when they are infected with baculovirus due to the activation of the polyhedrin promoter/eGFP complex by baculovirus gene products expressed during the infection. Using a 96-well plate format end-point dilution assay, baculovirus titers can be determined in three days using a fluorescence microscope.