Set of Vectors for the Expression of Histidine-Tagged Proteins in Vaccinia Virus Recombinants. (March 2001)

• Record Type: Journal Article
• Authors: Galindo, Inmaculada; Lorenzo, María M.; Blasco, Rafael
• Is Part Of: Biotechniques. Volume 30:Number 3(2001)
• Page Start: 524
• Page End: 529
• Publication Date: 2001-03
• Subjects: Biology, Experimental -- Periodicals; Molecular biology -- Periodicals; Medical technology -- Periodicals; Biology, Experimental; Medical technology; Molecular biology; Clinical Laboratory Techniques -- Periodicals; Research -- Periodicals; Medical Laboratory Science -- Periodicals; Periodicals; Electronic journals; 570
• DOI: 10.2144/01303st03
• Languages: English
• ISSNs: 0736-6205

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.