Wang Qiang, Zhou Yang, Chen Keping, Ju Xiaoli
Institute of Life Sciences, Jiangsu University, Zhenjiang, Jiangsu, 212013, People's Republic of China.
School of Medicine, Jiangsu University, Zhenjiang, Jiangsu, 212013, People's Republic of China.
Mol Biotechnol. 2016 May;58(5):319-27. doi: 10.1007/s12033-016-9931-4.
Baculovirus expression vector systems (BEVSs) have been widely used for recombinant protein production. Many studies have tried to improve the recombinant protein production by either modification of BEVSs vectors or by cell line selection. In this study, using a modified shRNA vector, we established a stable Bombyx mori cell line that significantly inhibits expression of caspase-1 after selection. We further compared cell proliferation and viability between caspase-1-suppressed cells and control cells, and found that there is no significant difference in these stable cell lines. We utilized these cell lines to analyze recombinant protein production after infection with recombinant baculovirus. We found that both intracellular and extracellular recombinant protein production significantly increased when expression of caspase-1 is inhibited in BmN cells. These data indicate that blocking the apoptotic pathway is a promising way to enhance recombinant protein production in BEVSs.
杆状病毒表达载体系统（BEVSs）已被广泛用于重组蛋白的生产。许多研究试图通过修饰BEVSs载体或选择细胞系来提高重组蛋白的产量。在本研究中，我们使用一种修饰的短发夹RNA（shRNA）载体，建立了一个稳定的家蚕细胞系，该细胞系在筛选后能显著抑制半胱天冬酶-1的表达。我们进一步比较了半胱天冬酶-1抑制细胞和对照细胞之间的细胞增殖和活力，发现这些稳定细胞系之间没有显著差异。我们利用这些细胞系分析重组杆状病毒感染后的重组蛋白生产情况。我们发现，当BmN细胞中半胱天冬酶-1的表达受到抑制时，细胞内和细胞外的重组蛋白产量均显著增加。这些数据表明，阻断凋亡途径是提高BEVSs中重组蛋白产量的一种有前景的方法。
