一种新的植物miRNA体内下调表达系统

摘要/Abstract

摘要： MicroRNA（miRNA）是一种内源性的21到24个核苷酸长度的小分子RNA，在植物发育过程中起重要作用。旨在建立一种新的植物miRNA下调表达系统，为miRNA的功能研究提供有力的工具。不同于原有targetmimicry的构建方法，以miRNA核心序列为基础，通过设计特定DNA引物，利用聚合酶链式反应（PCR）获得多拷贝的靶基因类似序列（Targetmimicryortargetmimics），构建targetmimicry载体，转化拟南芥。通过qRT-PCR验证miRNA靶基因在转基因植株中的表达。以拟南芥6个保守miRNA为基础，获得了各个miRNA下调表达转基因株系，在转基因植株中检测miRNA靶基因的表达，与野生型相比差异明显，均发生了明显的上调。我们基于一种新的targetmimicry的载体构建方法，成功建立了一种在植物中具有普遍适用性的高效miRNA低表达系统。

关键词: miRNA, 低表达, 植物, 新方法

Abstract: MiRNA is a kind of endogenous small RNA about 21-25 nuleotides in length that plays important roles in plant morphology and development. The present study was performed to establish a novel system for plant miRNA knockdown in vivo, supply a enegetic tool for miRNA function analysis. Unlike the original target mimicry vector method, we obtained multi-copyed target mimicry sequences using self-ligation method with designed specific DNA fragment based on miRNA motif, construct expression vector and transform to Arabidopsis thaliana. Starting with the conserved miRNA in Arabidopsis, we obtained six transgene lines of miRNA knockdown. We detected the relative expression of related miRNA targets in transgene plants, resulting obviously up-regulated compared with the relative expression in wild type. Based on target mimicry vector construction, we obtained MIM stable lines in Arabidopsis, successfully established a general and high effective miRNA knockdown system in plant species.

Key words: miRNA , Knockdown, Plant , Novel method

姜锋,许硕,胡正,姜奇彦,张辉. 一种新的植物miRNA体内下调表达系统[J]. 生物技术通报, 2014, 0(9): 65-71.

Jiang Feng, Xu Shuo, Hu Zheng,Jiang Qiyan, Zhang Hui. A Novel Plant miRNA Knockdown System in Vivo[J]. Biotechnology Bulletin, 2014, 0(9): 65-71.

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