猪STC-1 siRNA设计及其基因沉默效率检测

Design and efficiency of gene silencing detection on pig STC-1 siRNA

根据猪肌肉斯钙素(STC)mRNA序列设计3对小干扰RNA(siRNA)、阴性对照siRNA,转染PK15细胞后,通过荧光观察转染效率和基因表达效率,利用实时荧光定量PCR(QRT-PCR)及Western blot测定细胞内siRNA基因沉默后STC-1siRNA及蛋白的表达水平。结果表明:3个STC-1siRNA转染细胞后均有较高的荧光信号;STC-132细胞组STC-1mRNA和STC-1蛋白相对表达水平都最低,该组与其他组差异极显著。结果说明成功设计出STC-1siRNA序列,为后续通过调控STC-1表达研究其在肌肉发育中的功能奠定了基础。

Three pairs of STC-1 siRNA and a pair of negative control siRNA were designed based on STC-1 mRNA sequence. After transfection of PK15 cell, efficiency of transfection and gene expression was observed by fluorescent tech- nique, and QRT-PCR and western blot were used to determine siRNA in cells that gene silencing on STC-1 siRNA and protein expression level. The results showed that three siRNA on STC-1 had a higher fluorescence signal after the trans fection cells; STC-1 mRNA and STC-1 protein expression levels were significantly lower in STC-132 cells than those in other groups （P〈0.01）. The study suggested that the successfully designed STC-1 siRNA sequences can provide a foun dation for subsequent control on STC-1 expression in the function of muscle development.