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一种用于快速简便病毒定量的新型转基因 Sf9 细胞系。

A Novel Transgenic Sf9 Cell Line for Quick and Easy Virus Quantification.

作者信息

Kim Kyu-Seek, Bae Jun-Su, Moon Hyuk-Jin, Kim Do-Young, Woo Soo-Dong

机构信息

Department of Agricultural Biology, College of Agriculture, Life & Environment Science, Chungbuk National University, Cheongju 28644, Republic of Korea.

IPBL Inc., Cheongju 28644, Republic of Korea.

出版信息

Insects. 2024 Sep 11;15(9):686. doi: 10.3390/insects15090686.

Abstract

The following study was conducted to generate a transgenic Sf9 cell line for rapid and easy virus quantification in the baculovirus expression system (BES). The hr3 (homologous region 3) and and promoters were used as the expression structures to induce rapid and intense expression of the enhanced green fluorescent protein gene in cells in response to viral infection. Of 20 transgenic Sf9 cell lines generated using the piggyBac system, the cell line that showed the highest fluorescence expression in the shortest time in response to viral infection was selected and named Sf9-QE. The average diameter of the Sf9-QE cells was around 16 μm, which is 2 μm smaller than the average diameter of Sf9 cells, whereas the rate of cell proliferation was around 1.6 times higher in the Sf9-QE cells. Virus quantification using the Sf9-QE cell line did not produce significantly different results compared to the other cell lines; however, the time required for complete virus quantification was approximately 5.3 to 6.0 days for the Sf9-QE cells, which is around 4 to 6 days shorter than the time required for the other cell lines, enabling convenient and accurate virus quantification via fluorescence photometry within around 6.0 to 6.3 days. The properties of the Sf9-QE cells were stable for up to at least 100 passages.

摘要

进行了以下研究,以生成一种转基因Sf9细胞系,用于杆状病毒表达系统(BES)中快速简便的病毒定量。hr3(同源区域3)和启动子用作表达结构,以在病毒感染时诱导细胞中增强型绿色荧光蛋白基因的快速强烈表达。在使用piggyBac系统产生的20个转基因Sf9细胞系中,选择了在病毒感染后最短时间内显示出最高荧光表达的细胞系,并命名为Sf9-QE。Sf9-QE细胞的平均直径约为16μm,比Sf9细胞的平均直径小2μm,而Sf9-QE细胞的细胞增殖率约高1.6倍。与其他细胞系相比,使用Sf9-QE细胞系进行病毒定量没有产生显著不同的结果;然而,Sf9-QE细胞完成病毒定量所需的时间约为5.3至6.0天,比其他细胞系所需的时间短约4至6天,从而能够在约6.0至6.3天内通过荧光光度法进行方便准确的病毒定量。Sf9-QE细胞的特性在至少100代内保持稳定。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f8e5/11431869/2523cffb1b50/insects-15-00686-g001.jpg

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