绵羊血管内皮生长因子基因siRNA载体在卵巢颗粒细胞中的表达

摘要

The aim of this research is to explore siRNA expression plasmid vector on gene VEGF,and the expression of this plasmid in granular cells of ovine ovary. The granular cells of ovine ovary were cultured in vitro using DMEM/F12 (containing 10％ FBS), and the four kinds of vectors constructed before were transfected into granular cells using lipofectamine 2000. The control group is granular cells transfected vector PGC, and the other three groups are granular cells transfected vectors PGC-1, PGC-2, PGC-3 respectively, named Group Ⅰ、 Group Ⅱ and Group Ⅲ. The expression of VEGF in fibroblast was calculated by standard curve which was drawn according to ELISA. Compared with the control,the expression of VEGF in treatment groups were partly lower 55.37％ ,81.45％ and 73.29％. The interference efficiency of PGC-2 is the best of all.%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.