TMV-Gate vectors: gateway compatible tobacco mosaic virus based expression vectors for functional analysis of proteins

• Published in: Scientific reports Vol. 2; no. 1; p. 874
• Main Authors: Kagale, Sateesh, Uzuhashi, Shihomi, Wigness, Merek, Bender, Tricia, Yang, Wen, Borhan, M Hossein, Rozwadowski, Kevin
• Format: Journal Article
• Language: English
• Published: England Nature Publishing Group 19.11.2012
• Subjects: Epitopes; Expression vectors; Genetic Vectors; Green Fluorescent Proteins - genetics; Plant Proteins - analysis; Plant Proteins - genetics; Plant Proteins - metabolism; Plant viruses; Plants - genetics; Plants - virology; Protein interaction; Protein purification; Proteins; Recombinant Proteins - biosynthesis; Tobacco Mosaic Virus - genetics
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/srep00874

Plant viral expression vectors are advantageous for high-throughput functional characterization studies of genes due to their capability for rapid, high-level transient expression of proteins. We have constructed a series of tobacco mosaic virus (TMV) based vectors that are compatible with Gateway technology to enable rapid assembly of expression constructs and exploitation of ORFeome collections. In addition to the potential of producing recombinant protein at grams per kilogram FW of leaf tissue, these vectors facilitate either N- or C-terminal fusions to a broad series of epitope tag(s) and fluorescent proteins. We demonstrate the utility of these vectors in affinity purification, immunodetection and subcellular localisation studies. We also apply the vectors to characterize protein-protein interactions and demonstrate their utility in screening plant pathogen effectors. Given its broad utility in defining protein properties, this vector series will serve as a useful resource to expedite gene characterization efforts.