High expression of foreign proteins from a biosafe viral vector derived from Turnip mosaic virus

A Touriño,F Ponz,A Fereres,F Sánchez

doi:10.5424/sjar/200806s1-373

Abstract

A vector derived from an original infectious clone of Turnip mosaic potyvirus (TuMV) has been developed. The vector (p35Tunos-vec01) was made through the creation of a new unique insertion site for foreign genes between the NIb and the CP genes of the viral genome. The jellyfish green fluorescent protein (GFP) and E. coli beta-glucuronidase (GUS) were expressed in Arabidopsis thaliana plants infected with chimeric vectors carrying their corresponding genes. In the case of GUS, expression levels that were 15-50 fold higher than nuclear transgenic A. thaliana lines carrying the same gene were attained. Acceptable levels of gene stability were found in the infected plants, compatible with the use of the vector for protein production in plants. A non-aphid transmissible version of the vector was also made and its lack of aphid transmission was extensively shown. The vector was not transmitted through seeds or by contact between plants.

Highlights

Altos niveles de expresión de proteínas foráneas a partir de un vector viral bioseguro derivado del virus del mosaico del nabo
We introduced this mutation in vec01 to produce vec01-NAT1 (Table 1) and tested infectivity, foreign gene expression, and aphid transmissibility
A viral vector derived from an infectious Turnip mosaic potyvirus (TuMV) clone

Summary

Introduction

Altos niveles de expresión de proteínas foráneas a partir de un vector viral bioseguro derivado del virus del mosaico del nabo. For about 20 years (French et al, 1986), different research groups have developed a range of virus-derived vectors which have been used to express a variety of gene products (Pogue et al, 2002; Grill et al, 2005). Some specific points such as foreign gene stability within the context of the viral genome and insert size are still an issue, virus-derived vectors are an important biotechnological tool for foreign gene expression, and their development is making progress. Members of the Potyviridae family have been found in practically all know taxonomical families of cultivated and non-cultivated plants Virions of this group of viruses are flexuous rods, made up of ca. Viral proteins mediating aphid transmission are the CP, involving its DAG triplet, and the helper component (HC-Pro), in which several residues have been revealed as essential for transmission (Maia and Bernardi, 1996; Maia et al, 1996)

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