黄瓜花叶病毒基因沉默载体的效率和稳定性分析

发布时间：2018-02-20 21:25

  本文关键词： 病毒诱导的基因沉默 黄瓜花叶病毒 b 八氢番茄红素脱氢酶　出处：《浙江农业学报》2017年04期 　论文类型：期刊论文

【摘要】：病毒诱导的基因沉默(virus-induced gene silencing,VIGS)已被广泛应用于植物基因的功能研究。试验以八氢番茄红素脱氢酶(phytoene desaturase,PDS)作为指示基因,在本生烟体内分析黄瓜花叶病毒(Cucumber mosaic virus,CMV)作为VIGS载体对植物内源PDS mRNA的沉默效率。采用分子克隆的方法,将FnyCMV 2b ORF替换为不同长度的PDS序列,并分析了其在本生烟植物上诱导的PDS沉默效率。结果表明,该重组病毒能诱导上部系统叶的光漂白表型,且沉默效率与插入片段长度呈一定的正相关性,但在顶端新生叶中均不引起明显的光漂白现象,过长的片段插入(≥518 nt)影响病毒基因组的遗传稳定性,导致插入片段的部分缺失。CMV作为VIGS载体,最适合的插入片段约为350 nt。
[Abstract]:Virus-induced gene silencing (VIGS) has been widely used to study the function of plant genes. Phytoene desaturase (PDS) was used as an indicator gene. The silencing efficiency of cucumber mosaic virus (Cucumber mosaic virusus) to plant endogenous PDS mRNA was analyzed in vivo. FnyCMV 2b ORF was replaced with different length PDS sequences by molecular cloning. The results showed that the recombinant virus could induce the photobleaching phenotype of the upper leaves, and the silencing efficiency was positively correlated with the length of the inserted fragment. However, there was no obvious photobleaching in the apical neonate leaves. The long fragment insertion (鈮，

本文编号：1520062