摘要
为在昆虫细胞中表达鸭源新城疫病毒NP蛋白质,首先根据鸭源新城疫病毒NP基因序列设计引物,扩增出NP基因,将其克隆至杆状病毒表达载体p Fast Bac1,获得重组转移载体p Fast Bac-NP,将其转化到DH10Bac感受态细胞中,经抗性和蓝白斑筛选,获得重组穿梭质粒r Bacmid-NP,在脂质体的介导下转染昆虫细胞Sf9,获得重组杆状病毒NP蛋白质。Western blot、间接免疫荧光试验结果均显示表达的重组蛋白质能够与全病毒阳性血清发生特异性反应,具有良好的反应原性。以上结果说明鸭源新城疫病毒NP蛋白质在昆虫细胞中获得了成功表达。
To express the NP gene of duck NDV ( Newcastle disease virus) in insect cells, one pair of specific prim?ers were designed for the amplification of NP gene by PCR, and the amplified fragment was cloned into baculovirus expres?sion vector pFastBac1. The recombinant vector pFastBac?NP was transformed into DH10Bac Escherichia coli, and the posi?tive recombinant bacmid rBacmid?NP was screened according to the resistance and the blue?white plague screening. The NP protein was aquired through the transfection of rBacmid?NP into Sf9 insect cells by liposome. The results of Western blot and indirect immunofluorescence showed that the recombinant proteins could be recognized by positive serum, indicating that the protein had good reactiongenicity. These results suggested that the NP proteins of duck NDV had been successfully expressed in insect cells.
出处
《江苏农业学报》
CSCD
北大核心
2015年第6期1384-1388,共5页
Jiangsu Journal of Agricultural Sciences
基金
国家自然科学基金项目(31302096)
江苏省农业科技支撑计划项目(BE2013415)
江苏省"六大人才高峰"项目(NY-009)
