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Expresión of functionally active helper component protein of Tobacco etch potyvirus in the yeast Pichia pastoris

AuthorsRuiz-Ferrer, Virginia; Goytia, Elisa; Martínez-García, Belén ; López-Abella, Dionisio; López-Moya Gómez, Juan José
Issue Date1-Jan-2004
PublisherSociety for General Microbiology
CitationJournal of General Virology, 85 (1) : 241-249
AbstractTobacco etch potyvirus (TEV) is transmitted by aphids in a non-persistent manner with the assistance of a virus-encoded protein known as helper component (HC-Pro). To produce a biologically active form of recombinant TEV HC-Pro protein, heterologous expression in the methylotrophic yeast Pichia pastoris was used. A cDNA encoding the TEV HC-Pro region, fused to a Saccharomyces cerevisiae α-mating factor secretory peptide coding region, was inserted into the P. pastoris genome using a modified version of the pPIC9 vector. The expressed TEV HC-Pro protein was obtained directly from culture medium of recombinant yeast colonies; it was able to interact with TEV particles in a protein overlay binding assay, and also to assist aphid transmission of purified TEV particles to plants using the aphid Myzus persicae as vector. Our results indicate that P. pastoris provides a rapid and low-cost heterologous expression system that can be used to obtain biologically active potyvirus HC-Pro protein for in vitro transmission assays
Description9 páginas, 3 figuras, 1 tabla -- PAGS NROS. 241-249
Publisher version (URL)http://dx.doi.org/10.1099/vir.0.19557-0
Appears in Collections:(CIB) Artículos
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