杆状病毒感染后 sf-caspase-1 抑制稳定细胞中增强的重组蛋白生产和分子伴侣的差异表达。
Enhanced recombinant protein production and differential expression of molecular chaperones in sf-caspase-1-repressed stable cells after baculovirus infection.
机构信息
Institute of Biotechnology, Department of Life Science, National Tsing Hua University, No. 101, Section 2, Kuang-Fu Road, Hsinchu 30013, Taiwan, R.O.C.
出版信息
BMC Biotechnol. 2012 Nov 7;12:83. doi: 10.1186/1472-6750-12-83.
BACKGROUND
There are few studies that have examined the potential of RNA inference (RNAi) to increase protein production in the baculovirus expression vector system (BEVS). Spodoptera frugiperda (fall armyworm) (Sf)-caspase-1-repressed stable cells exhibit resistance to apoptosis and enhancement of recombinant protein production. However, the mechanism of recombinant protein augmentation in baculovirus-infected Caspase-repressed insect cells has not been elucidated.
RESULTS
In the current study, we utilized RNAi-mediated Sf-caspase-1-repressed stable cells to clarify how the resistance to apoptosis can enhance both intracellular (firefly luciferase) and extracellular (secreted alkaline phosphatase [SEAP]) recombinant protein production in BEVS. Since the expression of molecular chaperones is strongly associated with the maximal production of exogenous proteins in BEVS, the differential expression of molecular chaperones in baculovirus-infected stable cells was also analyzed in this study.
CONCLUSION
The data indicated that the retention of expression of molecular chaperones in baculovirus-infected Sf-caspase-1-repressed stable cells give the higher recombinant protein accumulation.
背景
目前鲜有研究探讨 RNA 干扰 (RNAi) 提高杆状病毒表达载体系统 (BEVS) 中蛋白产量的潜力。秋粘虫 (Sf)-caspase-1 抑制稳定细胞表现出抗凋亡和增强重组蛋白产量的特性。然而，杆状病毒感染 Caspase 抑制昆虫细胞中重组蛋白增加的机制尚未阐明。
结果
本研究利用 RNAi 介导的 Sf-caspase-1 抑制稳定细胞，阐明了抗凋亡如何增强 BEVS 中细胞内（萤火虫荧光素酶）和细胞外（分泌碱性磷酸酶 [SEAP]）重组蛋白的产量。由于分子伴侣的表达与 BEVS 中外源蛋白的最大产量密切相关，因此本研究还分析了杆状病毒感染稳定细胞中分子伴侣的差异表达。
结论
数据表明，杆状病毒感染 Sf-caspase-1 抑制稳定细胞中分子伴侣的表达保留导致更高的重组蛋白积累。
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