@article{mbs:/content/journal/jgv/10.1099/vir.0.19557-0, author = "Ruiz-Ferrer, Virginia and Goytia, Elisa and Martínez-García, Belén and López-Abella, Dionisio and López-Moya, Juan José", title = "Expression of functionally active helper component protein of Tobacco etch potyvirus in the yeast Pichia pastoris", journal= "Journal of General Virology", year = "2004", volume = "85", number = "1", pages = "241-249", doi = "https://doi.org/10.1099/vir.0.19557-0", url = "https://www.microbiologyresearch.org/content/journal/jgv/10.1099/vir.0.19557-0", publisher = "Microbiology Society", issn = "1465-2099", type = "Journal Article", abstract = " Tobacco 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.", }