hpRNA的制备新方法及其在RNAi载体中的应用
A Prepared Method of hpRNA and Its Application for RNAi Vector
刘晓宁1
作者信息
- 1. 黄河科技学院医学院,河南郑州450063
- 折叠
摘要
[目的]验证制备新方法所获得的稳定hpRNA是否可用于RNAi载体的构建并达到基因表达干扰的效果.[方法]利用制备新方法(2μL DNA与8μL H2O混匀,95℃ 5 min,37 ℃ 15 min)制备hpRNA,后采用REGS方法构建ZmMDAR RNAi载体,通过Grx1-roGFP2和荧光实时定量PCR检测基因表达干扰的效果.[结果]利用制备新方法制备50 nt hpRNA.ZmMDAR基因的表达量变化和Grx1-roGFP2探针的荧光比值变化表明hpRNA介导的干扰载体引起了原生质体氧化还原状态的变化.[结论]通过制备新方法所制备的hpRNA为RNAi载体所介导的干扰作用奠定基础.
Abstract
[Objective] The aim was to test a method if it is applied for the RNAi vector and lead to interference effect.[Method] We prepared hpRNA by the new method (2 μL DNA and 8 μL H2O mixed,95 ℃,5 min,37 ℃,15 min),constructed ZmMDAR RNAi vector with REGS method,and tested the interference effect of gene expression by Grxl-roGFP2 and reall-time PCR.[Result] Using new method for preparing 50nt hpRNA,change of Zm MDAR expression and fluorescence ratio of Grxl-roGFP2 indicated that hpRNA mediated interference vector caused the change of redox state of protoplast.[Conclusion] The prepared hpRNA by new method will lay a foundation for interference effect mediated by RNAi vector.
关键词
发夹/RNA/REGS/ZmMDAR/RNAi/Grx1-roGFP2Key words
hpRNA/REGS/ZmMDAR RNAi/Grxl-roGFP2引用本文复制引用
出版年
2016
国家科技期刊平台
NSTL
维普
万方数据