Set of vectors for the expression of histidine-tagged proteins in vaccinia virus recombinants |
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Authors: | Galindo I Lorenzo M M Blasco R |
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Institution: | Departamento de Mejora genética y biotecnología, INIA, Ctra. La Coru?a km 7, E-28040 Madrid, Spain. |
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Abstract: | Vaccinia virus expression vectors are widely used to direct the expression of proteins in eukaryotic cells. Here, we describe a new set of plasmid vectors designed for the expression of histidine-tagged proteins in the vaccinia system. To facilitate the rapid isolation of virus recombinants, the plasmids contain a viral gene (F13L) that serves as an efficient selection marker based on virus plaque phenotype. Histidine codons and restriction sites derived from pET-16b bacterial expression plasmid were included, thus facilitating the transfer of genes between E. coli and vaccinia expression plasmids. Plasmids in which the gene is placed downstream of either a strong vaccinia virus or a T7 promoter were constructed, allowing for constitutive or conditional expression, respectively, of the foreign protein. |
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