Efficiency of VIGS and gene expression in a novel bipartite potexvirus vector delivery system as a function of strength of TGB1 silencing suppression.

Abstract

We have developed plant virus-based vectors for virus-induced gene silencing (VIGS) and protein expression, based on Alternanthera mosaic virus (AltMV), for infection of a wide range of host plants including Nicotiana benthamiana and Arabidopsis thaliana by either mechanical inoculation of in vitro transcripts or via agroinfiltration. In vivo transcripts produced by co-agroinfiltration of bacteriophage T7 RNA polymerase resulted in T7-driven AltMV infection from a binary vector in the absence of the Cauliflower mosaic virus 35S promoter. An artificial bipartite viral vector delivery system was created by separating the AltMV RNA-dependent RNA polymerase and Triple Gene Block (TGB)123-Coat protein (CP) coding regions into two constructs each bearing the AltMV 5' and 3' non-coding regions, which recombined in planta to generate a full-length AltMV genome. Substitution of TGB1 L(88)P, and equivalent changes in other potexvirus TGB1 proteins, affected RNA silencing suppression efficacy and suitability of the vectors from protein expression to VIGS.

• Authors: Hyoun-Sub Lim, Anna Maria Vaira, Leslie L Domier, Sung Chul Lee, Hong Gi Kim, John Hammond
• Journal: Virology (Virology) Vol. 402 Issue 1 Pg. 149-63 (Jun 20 2010) ISSN: 1096-0341 [Electronic] United States
• PMID: 20381827 (Publication Type: Journal Article)
• Copyright: Published by Elsevier Inc.

Chemical References

• RNA-Binding Proteins

Topics

• Amino Acid Substitution (genetics)
• Arabidopsis (virology)
• Bacteriophage T7 (genetics)
• Caulimovirus (genetics)
• Gene Expression
• Gene Silencing
• Genes, Viral
• Genetic Vectors
• Mutation, Missense
• Potexvirus (genetics, immunology)
• Promoter Regions, Genetic
• RNA-Binding Proteins (genetics)
• Tobacco (virology)