A novel strategy for the expression of foreign genes from plant virus vectors

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	A novel strategy for the expression of foreign genes from plant virus vectors
	Toth, R. L.
刊名	Febs Letters
	2001
卷号	489 期号:42038 页码:215-219
ISSN号	0014-5793
中文摘要	Potato virus X (PVX)-based vector constructs were generated to investigate the use of an internal ribosome entry site (IRES) sequence to direct translation of a viral gene. The 148-nucleotide IRESep sequence from a crucifer-infecting strain of tobacco mosaic virus was used to direct expression of the PVX coat protein (CF), The IRES,vas inserted downstream of the gene encoding green fluorescent protein (GFP) and upstream of the PVX CP, in either sense or antisense orientation, such that CP expression depended on ribosome recruitment to the IRES. Stem-loop structures were inserted at either the 3 '- or 5 ' -end of the IRES sequence to investigate its mode of action. In vitro RNA transcripts were inoculated to Nicotiana benthamiana plants and protoplasts: levels of GFP and CP expression were analysed by enzyme-linked immunosorbent assay and the rate of virus cell-to-cell movement was determined by confocal laser scanning microscope imaging of GFP expression, PVX CP was expressed, allowing cell-to-cell movement of virus, from constructs containing the IRES sequence in either orientation, and from the construct containing a stem-loop structure at the 5 ' -end of the IRES sequence, No CP was expressed from a construct containing a stem-loop at the 3 ' -end of the IRES sequence. Our results suggest that the IRES sequence is acting in vivo to direct expression of the 3 ' -proximal open reading frame in a bicistronic mRNA thereby demonstrating the potential of employing IRES sequences for the expression of foreign proteins from plant virus-based vectors. (C) 2001 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved.
语种	英语
公开日期	2015-06-08
内容类型	期刊论文
源URL	[http://ir.xtbg.org.cn/handle/353005/8755]
专题	西双版纳热带植物园_公共技术服务中心主要仪器相关文献_激光共聚焦显微镜
推荐引用方式 GB/T 7714	Toth, R. L.. A novel strategy for the expression of foreign genes from plant virus vectors[J]. Febs Letters,2001,489(42038):215-219.
APA	Toth, R. L..(2001).A novel strategy for the expression of foreign genes from plant virus vectors.Febs Letters,489(42038),215-219.
MLA	Toth, R. L.."A novel strategy for the expression of foreign genes from plant virus vectors".Febs Letters 489.42038(2001):215-219.