摘要
旨在评价H1N1猪流感病毒感染猪血管内皮细胞(PUVEC)后,肽基脯氨酰异构酶A(Ppia)、β-肌动蛋白(Actb)、核糖体蛋白L4(Rpl4)、酪氨酸3/色氨酸5-单加氧酶激活蛋白(Ywhaz)和甘油醛-3-磷酸脱氢酶(Gapdh)等5个内参基因的稳定性.采用Real-time RT-PCR方法测定H1N1 SIV感染PUVEC后3、6、9、12、15、18、24和30h时5个内参基因mRNA的表达水平,未感染PUVEC为对照,采用2-△Ct值对内参基因进行相对定量,应用Genorm软件对其稳定性进行评价.结果表明,5个内参基因的稳定性由高到低分别为Ppia和Rpl4、Ywhaz、Actb、Gapdh,其中Ppia和Rp14稳定性相同.据此可知,Ppia和Rpl4在所选的5个内参基因中是研究H1N1 SIV与机体互作理想的2个内参基因.
The purpose of this study is to analysis the expression stability of Ppia, Actb, Rpl4, Ywhaz and Gapdh genes in porcine umbilical vein endothelial cells infected by H1N1 swine influenza virus. The paper detected mRNA expression level of these five genes at 3h, 6h, 9h, 12h,15h,18h, 24h and 30h post-infection using real-time RT-PCR, and the uninfected group was set as a control group. Relative quantification of these 5 genes was calculated using 2^-△Ct value and then analysis of their expression stabilities was conducted using Genorm soffwere. The result showed that the expression stability of these 5 genes ranges in a decreasing order as follows： Ppia, Rpl4, Ywhaz, Actb, and Gapdh, among which Ppia and Rpl4 have the same expression stability. In sum, the Ppia and Rp14 genes are the ideal reference genes in H1N1 SIV infected PUVEC.
出处
《西南民族大学学报（自然科学版）》
CAS
2014年第4期489-494,共6页
Journal of Southwest Minzu University(Natural Science Edition)
基金
国家自然科学基金项目(编号31172307)
