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Title 

Development of tobacco ringspot virus-based vectors for foreign gene expression and virus-induced gene silencing in a variety of plants

Authors 

F ZhaoSeungmo LimD IgoriRan Hee YooSuk Yoon KwonJae Sun Moon

Publisher 

Elsevier

Issue Date 

2016

Citation 

Virology, vol. 492, no. 0, pp. 166-178

Keywords 

ArabidopsisCucurbitsForeign gene expressionFunctional genomicsLegumesNicotiana benthamianaPhytoene desaturaseTobacco ringspot virusVirus-induced gene silencing

Abstract 

We report here the development of tobacco ringspot virus (TRSV)-based vectors for the transient expression of foreign genes and for the analysis of endogenous gene function in plants using virus-induced gene silencing. The jellyfish green fluorescent protein (GFP) gene was inserted between the TRSV movement protein (MP) and coat protein (CP) regions, resulting in high in-frame expression of the RNA2-encoded viral polyprotein. GFP was released from the polyprotein via an N-terminal homologous MP-CP cleavage site and a C-terminal foot-and-mouth disease virus (FMDV) 2 A catalytic peptide in Nicotiana benthamiana. The VIGS target gene was introduced in the sense and antisense orientations into a SnaBI site, which was created by mutating the sequence following the CP stop codon. VIGS of phytoene desaturase (PDS) in N. benthamiana, Arabidopsis ecotype Col-0, cucurbits and legumes led to obvious photo-bleaching phenotypes. A significant reduction in PDS mRNA levels in silenced plants was confirmed by semi-quantitative RT-PCR.

ISSN 

0042-6822

Link 

http://dx.doi.org/10.1016/j.virol.2016.02.025

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2019-05-02


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