摘要
研究RNA干扰(RNA interference,RNAi)表达载体对UCP基因的抑制作用。针对鸡解耦联蛋白(avianuncoupling protein,av UCP)基因,构建4种RNAi质粒表达载体(分别称为A、B、C和D号载体)。再经过脂质体法转染成肌纤维细胞,应用荧光定量PCR及流式细胞术检测其对av UCP mRNA及蛋白表达的影响。构建的4种表达载体均抑制了av UCP的mRNA及蛋白的表达,其中转染C号载体的成肌纤维细胞av UCPmRNA表达相当于空白对照组的14.6%,蛋白抑制率达93.7%。RNAi表达载体可以有效的抑制avUCP基因的表达。
To investigate the inhibitory effects of RNAi（RNA interference） expression vector on the expression of a＇oUCP gene. Four siRNA expression vectors （A, B, C and D） were constructed to be aimed directly at avUCP gene. The recombinants were transfected into myofihrohlast with liposomes. Expression of avUCP was detected with fluorescence quantitative PCR （FQ- PCR）and flow cytometry. Four kinds of expression vectors could reduce the expression of avUCP mRNA and protein all in myofihroblast. The expression of avUCP mRNA reduced to 14. 6% of control group, and the protein inhibition ratio reached 93.7%. The RNAi expression vector can effectively inhibit the expression of avUCP gene.
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2007年第4期352-355,共4页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
"863"计划资助项目(2003AA243046)
"973"计划资助项目(2004CB117507)
关键词
RNA干扰
avUCP
表达载体
FQ-PCR
成肌纤维
RNA interference （RNAi）
avUCP
expression vector
fluorescence quantitative PCR （FQ-PCR）
myofihroblast
