Get 20M+ Full-Text Papers For Less Than $1.50/day. Start a 14-Day Trial for You or Your Team.

Learn More →

Retracted: An enhanced transient expression system in plants based on suppression of gene silencing by the p19 protein of tomato bushy stunt virus

Retracted: An enhanced transient expression system in plants based on suppression of gene... Transient gene expression is a fast, flexible and reproducible approach to high‐level expression of useful proteins. In plants, recombinant strains of Agrobacterium tumefaciens can be used for transient expression of genes that have been inserted into the T‐DNA region of the bacterial Ti plasmid. A bacterial culture is vacuum‐infiltrated into leaves, and upon T‐DNA transfer, there is ectopic expression of the gene of interest in the plant cells. However, the utility of the system is limited because the ectopic protein expression ceases after 2–3 days. Here, we show that post‐transcriptional gene silencing (PTGS) is a major cause for this lack of efficiency. We describe a system based on co‐expression of a viral‐encoded suppressor of gene silencing, the p19 protein of tomato bushy stunt virus (TBSV), that prevents the onset of PTGS in the infiltrated tissues and allows high level of transient expression. Expression of a range of proteins was enhanced 50‐folds or more in the presence of p19 so that protein purification could be achieved from as little as 100 mg of infiltrated leaf material. The effect of p19 was not saturated in cells that had received up to four individual T‐DNAs and persisted until leaf senescence. Because of its simplicity and rapidity, we anticipate that the p19‐enhanced expression system will have value in industrial production as well as a research tool for isolation and biochemical characterisation of a broad range of proteins without the need for the time‐consuming regeneration of stably transformed plants. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png The Plant Journal Wiley

Retracted: An enhanced transient expression system in plants based on suppression of gene silencing by the p19 protein of tomato bushy stunt virus

Download PDF
8 pages

Loading next page...
 
/lp/wiley/retracted-an-enhanced-transient-expression-system-in-plants-based-on-3dj6OuXEYt

References (40)

Publisher
Wiley
Copyright
Copyright © 2003 Wiley Subscription Services, Inc., A Wiley Company
ISSN
0960-7412
eISSN
1365-313X
DOI
10.1046/j.1365-313X.2003.01676.x
Publisher site
See Article on Publisher Site

Abstract

Transient gene expression is a fast, flexible and reproducible approach to high‐level expression of useful proteins. In plants, recombinant strains of Agrobacterium tumefaciens can be used for transient expression of genes that have been inserted into the T‐DNA region of the bacterial Ti plasmid. A bacterial culture is vacuum‐infiltrated into leaves, and upon T‐DNA transfer, there is ectopic expression of the gene of interest in the plant cells. However, the utility of the system is limited because the ectopic protein expression ceases after 2–3 days. Here, we show that post‐transcriptional gene silencing (PTGS) is a major cause for this lack of efficiency. We describe a system based on co‐expression of a viral‐encoded suppressor of gene silencing, the p19 protein of tomato bushy stunt virus (TBSV), that prevents the onset of PTGS in the infiltrated tissues and allows high level of transient expression. Expression of a range of proteins was enhanced 50‐folds or more in the presence of p19 so that protein purification could be achieved from as little as 100 mg of infiltrated leaf material. The effect of p19 was not saturated in cells that had received up to four individual T‐DNAs and persisted until leaf senescence. Because of its simplicity and rapidity, we anticipate that the p19‐enhanced expression system will have value in industrial production as well as a research tool for isolation and biochemical characterisation of a broad range of proteins without the need for the time‐consuming regeneration of stably transformed plants.

Journal

The Plant Journal – Wiley

Published: Mar 1, 2003

Keywords: ; ; ; ;

There are no references for this article.