利用“一步法”菜豆斑驳病毒（BPMV）病毒载体在豌豆（Pisum sativum L.）中进行病毒诱导的基因沉默（VIGS）和外源基因表达

Virus-Induced Gene Silencing (VIGS) and Foreign Gene Expression in Pisum sativum L. Using the "One-Step" Bean pod mottle virus (BPMV) Viral Vector.

作者信息

Meziadi Chouaïb, Blanchet Sophie, Geffroy Valérie, Pflieger Stéphanie

机构信息

Institute of Plant Sciences-Paris Saclay (IPS2), CNRS, INRA, Université Paris-Sud, Université d'Evry, Université Paris Saclay, Bâtiment 630, rue Noetzlin, CS 800004, 91192, Gif sur Yvette CEDEX, France.

Institute of Plant Sciences-Paris Saclay (IPS2), Université Paris Diderot, Sorbonne Paris-Cité, Bâtiment 630, rue Noetzlin, CS 80004, 91192, Gif sur Yvette CEDEX, France.

出版信息

Methods Mol Biol. 2017;1654:311-319. doi: 10.1007/978-1-4939-7231-9_23.

DOI:10.1007/978-1-4939-7231-9_23

PMID:28986801

Abstract

Plant viral vectors have been developed to facilitate gene function studies especially in plant species not amenable to traditional mutational or transgenic modifications. In the Fabaceae plant family, the most widely used viral vector is derived from Bean pod mottle virus (BPMV). Originally developed for overexpression of foreign proteins and VIGS studies in soybean, we adapted the BPMV-derived vector for use in other legume species such as Phaseolus vulgaris and Pisum sativum. Here, we describe a protocol for efficient protein expression and virus-induced gene silencing (VIGS) in Pisum sativum leaves and roots using the "one-step" Bean pod mottle virus (BPMV) viral vector.

摘要

植物病毒载体已被开发出来，以促进基因功能研究，特别是在不适合传统突变或转基因修饰的植物物种中。在豆科植物家族中，使用最广泛的病毒载体源自菜豆荚斑驳病毒（BPMV）。最初开发该载体是用于大豆中外源蛋白的过表达和病毒诱导的基因沉默（VIGS）研究，我们对源自BPMV的载体进行了改造，使其可用于其他豆科物种，如菜豆和豌豆。在此，我们描述了一种使用“一步法”菜豆荚斑驳病毒（BPMV）病毒载体在豌豆叶片和根中进行高效蛋白表达和病毒诱导基因沉默（VIGS）的方案。