BioTechniques (Mar 2001)

Set of Vectors for the Expression of Histidine-Tagged Proteins in Vaccinia Virus Recombinants

  • Inmaculada Galindo,
  • María M. Lorenzo,
  • Rafael Blasco

DOI
https://doi.org/10.2144/01303st03
Journal volume & issue
Vol. 30, no. 3
pp. 524 – 529

Abstract

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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.