Effects of seven RNA silencing suppressors on heterologous expression of green fluorescence protein expression mediated by a plant virus-based system in Nicotiana benthamiana.
- Author:
Sheng WANG
1
;
Jie DONG
;
Min CAO
;
Hongzhen MU
;
Guoping DING
;
Hong ZHANG
Author Information
- Publication Type:Journal Article
- MeSH: Genetic Vectors; Green Fluorescent Proteins; genetics; Plant Viruses; genetics; Plants, Genetically Modified; genetics; metabolism; Potexvirus; genetics; RNA Interference; Tobacco; genetics; metabolism
- From: Journal of Southern Medical University 2012;32(11):1536-1542
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo test the effects of 7 virus-encoded RNA silencing suppressors (RSSs) for enhancement of a plant virus-based vector system-mediated heterologous expression of green fluorescence protein (GFP) in Nicotiana benthamiana.
METHODSSeven transient expression vectors for the 7 RSSs were constructed and co-inoculated on the leaves of Nicotiana benthamiana with PVXdt-GFP vector, a novel Potato virus X-based plant expression vector, through agroinfiltration. The protein and mRNA expression levels of the reporter gene GFP in the co-inoculated Nicotiana leaves were examined by Western blotting, ELISA and RT-qPCR to assess the effect of the RSSs for GFP expression enhancement.
RESULTSThe 7 RSSs differed in the degree and duration of enhancement of heterologous GFP expression, and the p19 protein of Tomato bushy stunt virus (TBSV) induced the highest expression of GFP. African cassava mosaic virus AC2 protein and Rice yellow mettle virus P1 protein produced no obvious enhancement GFP expression.
CONCLUSIONTransient co-expression of RSSs suppresses host silencing response to allow high-level and long-term expression of heterologous genes in plant, but the optimal RSS has to be identified for each plant virus-based expression vector system.
