经密码子优化的猪Ficolin α在昆虫细胞的表达及其体外抗病毒作用初步分析

Expression of Codon-optimized Porcine Ficolinαin Insect Cell and Its Antiviral Activity in vitro

利用Bac-to-Bac昆虫杆状病毒表达系统表达具有天然构象的猪Ficolinα。首先构建重组穿梭质粒rBacmid-Ficolinα,转染sf9细胞获得重组杆状病毒rBV-Ficolinα;通过Image J软件对优化表达的Western blot图片进行灰度分析确定较佳表达条件;纯化目的蛋白质后,进一步评价其体外抗病毒活性。Western blot结果显示以7.5MOI接种量感染的sf9细胞在96h时获得了较高的表达,同时获得的重组猪Ficolinα具有抗猪繁殖与呼吸综合征病毒(PRRSV)活性。猪Ficolinα在sf9昆虫细胞中获得成功表达,且具有良好的抗PRRSV活性。本研究可以为猪Ficolinα的抗病毒活性及作用机制研究提供物质基础。

To get native porcine Ficolin α, Bac-to-Bac baculovirus expression system was used. Firstly, rBacmid-Ficolin α was constructed, and then transfected into sf9 insect cells to obtain recombinant baculovirus rBV-Ficolin α. The optimized expression conditions were detected by Western blot and analyzed by gray level using Image J software. Followed by purification, antiviral activity of target protein was assessed in vitro. Western blot results showed that the expression level of Ficolin α in transfected sf 9 insect cells was higher at the infection （MOI） of 7.5 when they were harvested at 96 h, and porcine Ficolin α had the antivirus activity against porcine reproductive and respiratory syndrome virus （PRRSV）. In conclusion, porcine Ficolin α was successfully expressed in sf9 insect cells and showed strongly anti-PRRSV activity. This study provided a material basis for the study of antiviral activity and its mechanism of porcine Ficolin α.