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

Departamento de Biología de Plantas, Centro de Investigaciones Biológicas, (CIB, CSIC), Ramiro de Maeztu 9, 28040 Madrid, Spain

Correspondence
Juan José López-Moya
jjlopez{at}cib.csic.es

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 -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.

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				E. Goytia, L. Fernandez-Calvino, B. Martinez-Garcia, D. Lopez-Abella, and J. J. Lopez-Moya Production of plum pox virus HC-Pro functionally active for aphid transmission in a transient-expression system. J. Gen. Virol., November 1, 2006; 87(Pt 11): 3413 - 3423. [Abstract] [Full Text] [PDF]

				V. Ruiz-Ferrer, J. Boskovic, C. Alfonso, G. Rivas, O. Llorca, D. Lopez-Abella, and J. J. Lopez-Moya Structural Analysis of Tobacco Etch Potyvirus HC-Pro Oligomers Involved in Aphid Transmission J. Virol., March 15, 2005; 79(6): 3758 - 3765. [Abstract] [Full Text] [PDF]