表达sFat-1基因慢病毒载体的构建及病毒生产

Construction of a lentiviral bector gor sFat-1 gene expression and production of lentivirals

为了建立有效地用于转基因动物生产的慢病毒表达系统,以线虫C.briggsae的ω-3脂肪酸去饱和酶基因(sFat-1)为目的基因构建了慢病毒表达载体,通过与慢病毒包装载体PLP1、PLP2和PLP/VSVG一起共转染293FT细胞系生产了慢病毒颗粒,并对其感染哺乳动物细胞NIH3T3细胞系的能力进行了检测。结果表明,该系统所生产的病毒滴度约为5.0×106TU/mL,这种病毒能够感染哺乳动物细胞,说明构建的慢病毒表达系统是有效的,应用其可以生产感染力良好的慢病毒颗粒,并高效地进行sFat-1基因转移。

In this research, a lentiviral vector system was constructed for the expression of sFat-1, a gene from C. briggsae encoding ω-3 fatty acid desaturase. By co-transfection of plasmids PLP1, PLP2, and PLP/VSVG with this vector, lentiviral particle produced 293FT cells, and the infection capacity of the lentiviral particle was tested by NIH3T3 cell line. Results demonstrated that this lentiviral expression system was successfully established,and it could be used to produce lentiviral particle with high efficient infection capacity for sFat-1 gene delivery.