title: Development of tobacco ringspot virus-based vectors for foreign gene expression and virus-induced gene silencing in a variety of plants.
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.
authors.affiliation: Biosystems and Bioengineering Program, University of Science and Technology (UST), Daejeon 305-350, Republic of Korea; Molecular Biofarming Research Center, Korea Research Institute of Bioscience & Biotechnology, Daejeon 305-806, Republic of Korea.
authors.forename: Fumei
authors.initials: F
authors.lastname: Zhao
authors.affiliation: Biosystems and Bioengineering Program, University of Science and Technology (UST), Daejeon 305-350, Republic of Korea; Molecular Biofarming Research Center, Korea Research Institute of Bioscience & Biotechnology, Daejeon 305-806, Republic of Korea.
authors.forename: Seungmo
authors.initials: S
authors.lastname: Lim
authors.affiliation: Biosystems and Bioengineering Program, University of Science and Technology (UST), Daejeon 305-350, Republic of Korea; Molecular Biofarming Research Center, Korea Research Institute of Bioscience & Biotechnology, Daejeon 305-806, Republic of Korea.
authors.forename: Davaajargal
authors.initials: D
authors.lastname: Igori
authors.affiliation: Biosystems and Bioengineering Program, University of Science and Technology (UST), Daejeon 305-350, Republic of Korea; Molecular Biofarming Research Center, Korea Research Institute of Bioscience & Biotechnology, Daejeon 305-806, Republic of Korea.
authors.forename: Ran Hee
authors.initials: RH
authors.lastname: Yoo
authors.affiliation: Biosystems and Bioengineering Program, University of Science and Technology (UST), Daejeon 305-350, Republic of Korea; Molecular Biofarming Research Center, Korea Research Institute of Bioscience & Biotechnology, Daejeon 305-806, Republic of Korea. Electronic address: sykwon@kribb.re.kr.
authors.forename: Suk-Yoon
authors.initials: SY
authors.lastname: Kwon
authors.affiliation: Biosystems and Bioengineering Program, University of Science and Technology (UST), Daejeon 305-350, Republic of Korea; Molecular Biofarming Research Center, Korea Research Institute of Bioscience & Biotechnology, Daejeon 305-806, Republic of Korea. Electronic address: jsmoon@kribb.re.kr.
authors.forename: Jae Sun
authors.initials: JS
authors.lastname: Moon
mesh_terms: D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism / Q000821:virology; D001483:Base Sequence; D036022:Capsid Proteins / Q000235:genetics / Q000378:metabolism; D028464:Cucurbita / Q000235:genetics / Q000378:metabolism / Q000821:virology; D007887:Fabaceae / Q000235:genetics / Q000378:metabolism / Q000821:virology; D029544:Foot-and-Mouth Disease Virus / Q000235:genetics / Q000378:metabolism; D020868:Gene Silencing; D005818:Genetic Engineering; D005822:Genetic Vectors / Q000737:chemistry / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D017860:Nepovirus / Q000235:genetics* / Q000378:metabolism; D010088:Oxidoreductases / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D010641:Phenotype; D038761:Photobleaching; D030821:Plants, Genetically Modified; D020815:Polyproteins / Q000235:genetics / Q000378:metabolism; D059748:Proteolysis; D012333:RNA, Messenger / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D014026:Nicotiana / Q000235:genetics* / Q000378:metabolism / Q000821:virology; D014170:Transformation, Genetic
chemical_list: D036022:Capsid Proteins; D020815:Polyproteins; D012333:RNA, Messenger; C485192:green fluorescent protein, Aequorea victoria; D049452:Green Fluorescent Proteins; D010088:Oxidoreductases; C022398:phytoene dehydrogenase
keywords: Arabidopsis; Cucurbits; Foreign gene expression; Functional genomics; Legumes; Nicotiana benthamiana; Phytoene desaturase; Tobacco ringspot virus; Virus-induced gene silencing