中国畜牧兽医2017，Vol.44Issue(2)：303-310,8.DOI:10.16431/j.cnki.1671-7236.2017.02.001
靶向TLR9基因siRNA的筛选及其对BHV-1增殖规律的影响
Screening of siRNA Targeted for TLR9 Gene and Its Effect on the Proliferation Profile of BHV-1
摘要
Abstract
The aim of this study was to investigate the role of Toll-like receptors 9 (TLR9) of the bovine embryonic tracheal (EBTr) cells in the innate immune response mediated by bovine herpesvirus type 1(BHV-1).Using small interfering RNA (siRNA) technology,three siRNA interference sequences target for TLR9 were designed and synthesized in this study.After siRNA-TLR9 interference,the expression levels of TLR9 gene were detected by Real-time PCR.After 48 h,the expression levels of TLR9 mRNA induced by siRNA-TLR9A,siRNA-TLR9B and siRNA-TLR9C were reduced to 60.90％,24.05％ and 40.75％,respectively.Comparing with the control group,the expression levels of TLR9 mRNA were significantly inhibited by siRNA-TLR9A fragments at 12 to 72 h (P＜0.05),and after transfecting the best fragments and infecting with BHV-1,the BHV-1 DNA copy numbers of siRNA-TLR9A group were lower than BHV-1 DNA of the control group at 6 to 72 h.The specific siRNA fragments target for TLR9 were successfully screened out in this test,and demonstrated that inhibition of TLR9 expressions could reduce the BHV-1 proliferation in EBTr cells.关键词
小分子干扰RNA/牛胚胎气管细胞/Toll样受体9/牛疱疹病毒Ⅰ型/实时荧光定量PCRKey words
siRNA/EBTr cells/TLR9/BHV-1/Real-time PCR分类
农业科技引用本文复制引用
呼凤丽,徐丽媛,张志飞,周伟光..靶向TLR9基因siRNA的筛选及其对BHV-1增殖规律的影响[J].中国畜牧兽医,2017,44(2):303-310,8.基金项目
国家自然科学基金项目(31260597) （31260597）
