Effect of temperature on recombinant protein expression in Semliki Forest virus infected mammalian cell lines growing in serum- free suspension cultures
Effect of temperature on recombinant protein expression in Semliki Forest virus infected mammalian cell lines growing in serum- free suspension cultures
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温度对在无血清悬浮培养物中生长的塞姆利基森林病毒感染的哺乳动物细胞系中重组蛋白表达的影响
DOI：
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发表时间：
1998
期刊：
影响因子：
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通讯作者：
K. Lundstrom
中科院分区：
文献类型：
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作者：
E. Schlaeger;K. Lundstrom
将萤火虫荧光素酶基因导入塞姆利基森林病毒（SFV）载体，制备高效价重组SFV颗粒。SFV广泛的宿主范围允许在无血清悬浮培养中生长的四种哺乳动物细胞系有效感染和高水平表达。孵育温度对重组蛋白的表达水平和表达时间有显著影响。例如，在感染后19小时收获时，在33°C孵育的啮齿动物BHK和CHO细胞系的荧光素酶活性明显高于37°C。在33°C下，随着感染后时间的延长，特异性表达水平增加了10-20倍，达到50小时。相比之下，在37°C培养的细胞感染后26小时，荧光素酶活性显著下降。温度对人细胞系HEK293荧光素酶表达有轻微影响，对亚克隆293（EBNA）没有影响。
The firefly luciferase gene was introduced into the Semliki Forest virus (SFV) vector and high titer recombinant SFV particles generated. The broad host range of SFV allowed efficient infection and high level expression of four mammalian cell lines growing in serum-free suspension cultures. The incubation temperature had dramatic effects on the level and duration of recombinant protein expression. For example, the luciferase activity was significantly higher in the rodent BHK and CHO cell lines incubated at 33 °C compared to 37 °C when harvested 19 h post-infection. At 33 °C the specific expression levels increased 10–20 fold during prolongation of the post-infection time up to 50 h. In contrast, a significant decrease in luciferase activity was observed from 26 h post-infection for cell cultures incubated at 37 °C. Only a slight temperature effect on luciferase expression was seen in the human cell line HEK293 and no effect was observed for the subclone293(EBNA).