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为构建绵羊血管内皮生长因子(VEGF)基因siRNA载体并对其在绵羊颗粒细胞中的表达,采用DMEM/F12培养液对绵羊卵巢颗粒细胞进行体外培养,采用脂质体转染法将干扰载体转染到颗粒细胞中,用ELISA试剂盒测定转染后各组颗粒细胞中VEGF的表达量。转染阳性对照载体PGC的颗粒细胞组为对照组,转染PGC-1、PGC-2和PGC-3的颗粒细胞组分别为试验组Ⅰ、试验组Ⅱ和试验组Ⅲ。与对照组相比,各试验组中VEGF的表达量分别降低了55.37%、81.45%和73.29%,其中载体PGC-2所转染的细胞中VEGF的表达量最少,说明在3个载体中,PGC-2对VEGF的基因沉默效果最好。
To construct a siRNA vector of sheep vascular endothelial growth factor (VEGF) gene and its expression in sheep granulosa cells, sheep ovarian granulosa cells were cultured in vitro with DMEM / F12 medium, and the interference vector was transfered by lipofection Dyed into granulosa cells, with the ELISA kit measured after transfection in each group of granulosa cells VEGF expression. The group of granulosa cells transfected with PGC-PGC-PGC-PGC-1, PGC-2 and PGC-3 were test group Ⅰ, test group Ⅱ and test group Ⅲ respectively. Compared with the control group, the expression of VEGF in each experimental group was reduced by 55.37%, 81.45% and 73.29%, respectively. The expression of VEGF in the transfected cells was the lowest, which indicated that among the three vectors, PGC-2 has the best effect on VEGF gene silencing.