利用豆荚斑纹病毒感染克隆对大豆(甘氨酸max)病毒诱导的基因沉默和瞬时基因表达

Q1 Agricultural and Biological Sciences

Current protocols in plant biology Pub Date : 2016-06-10 DOI:10.1002/cppb.20012

Steven A. Whitham, Lori M. Lincoln, R. V. Chowda-Reddy, Jaime D. Dittman, Jamie A. O'Rourke, Michelle A. Graham

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引用次数: 12

摘要

病毒诱导基因沉默(VIGS)是测定植物基因功能的一种强大而快速的方法。VIGS的基础是，病毒基因组经过改造，使其能够携带植物基因片段，通常在200到300个碱基对大小范围内。重组病毒被用来感染实验植物，无论病毒侵入何处，靶基因或靶基因都会被沉默。因此，VIGS是短暂的，在几周的时间内，可以设计VIGS结构，然后通过RNA沉默靶基因产生功能丧失表型。在大豆(Glycine max)中，豆荚斑驳病毒(Bean pod mottle virus, BPMV)已被设计成沉默多种功能基因和外源基因过表达的有价值的工具。本方案描述了一种设计BPMV结构体并利用它们沉默或瞬时表达大豆基因的方法。©2016 by John Wiley &儿子,Inc。

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Virus-Induced Gene Silencing and Transient Gene Expression in Soybean (Glycine max) Using Bean Pod Mottle Virus Infectious Clones

Virus-induced gene silencing (VIGS) is a powerful and rapid approach for determining the functions of plant genes. The basis of VIGS is that a viral genome is engineered so that it can carry fragments of plant genes, typically in the 200 to 300 base pair size range. The recombinant viruses are used to infect experimental plants, and wherever the virus invades, the target gene or genes will be silenced. VIGS is thus transient, and in the span of a few weeks, it is possible to design VIGS constructs and then generate loss-of-function phenotypes through RNA silencing of the target genes. In soybean (Glycine max), the Bean pod mottle virus (BPMV) has been engineered to be valuable tool for silencing genes with diverse functions and also for over-expression of foreign genes. This protocol describes a method for designing BPMV constructs and using them to silence or transiently express genes in soybean. © 2016 by John Wiley & Sons, Inc.

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来源期刊

Current protocols in plant biology Agricultural and Biological Sciences-Plant Science

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期刊介绍： Sound and reproducible laboratory methods are the foundation of scientific discovery. Yet nuances that are critical for an experiment''s success are not captured in the primary literature but exist only as part of a lab''s oral tradition. Current Protocols in Plant Biology provides reproducible step-by-step instructions for protocols relevant to plant research. Furthermore, Current Protocols content is thoughtfully organized by topic for optimal usage and to maximize contextual knowledge. Quarterly issues allow Current Protocols in Plant Biology to constantly evolve to keep pace with the newest discoveries and developments. Current Protocols in Plant Biology is the comprehensive source for protocols in the multidisciplinary field of plant biology, providing an extensive range of protocols from basic to cutting edge. Coverage includes: Extraction and analysis of DNA, RNA, proteins Chromosome analysis Transcriptional analysis Protein expression Metabolites Plant enzymology Epigenetics Plant genetic transformation Mutagenesis Arabidopsis, Maize, Poplar, Rice, and Soybean, and more.