一种使用 Sf-9 Easy Titer 细胞系滴定杆状病毒库存的快速方法。
A rapid method for titrating baculovirus stocks using the Sf-9 Easy Titer cell line.
机构信息
Protein Expression Laboratory, Advanced Technology Program, SAIC-Frederick, National Cancer Institute at Frederick, Frederick, MD 21702, USA.
出版信息
Biotechniques. 2009 Sep;47(3):785-8. doi: 10.2144/000113238.
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.
已经开发出一种新的快速方法来滴定杆状病毒株，使用新型细胞系 Sf-9 Easy Titer（Sf-9ET）。Sf-9ET 细胞系已被转染含有增强型绿色荧光蛋白（eGFP）基因的质粒 DNA，该基因受杆状病毒多角体启动子的控制。在滴定测定中，当 Sf-9ET 细胞被杆状病毒感染时，由于杆状病毒基因产物在感染过程中表达而激活多角体启动子/eGFP 复合物，因此细胞会变成绿色。使用 96 孔板格式终点稀释测定法，使用荧光显微镜可以在三天内确定杆状病毒滴度。
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