Vaccinia virus, recombinant protein expression

MacDonald, M.R., Takeda, J., Rice, C.M. etal. (1989). Multiple tachykinins are produced and secreted upon post-translational processing of the three substance P precursor proteins, alpha-, beta-, and gamma-preprotachykinin. Expression of the preprotachykinins in AtT-20 cells infected with vaccinia virus recombinants. J. Biol. Chem. 264, 15578-15592. 

Janknecht, R., de Martynoff, G., Lou, J., Hipskind, R., Noerdheim, A. and Stunnen-berg, H. (1991) Rapid and efficient purification of native histidine-tagged protein expressed by recombinant vaccinia virus. Proc. Natl. Acad. Sci. USA, 88, 8972-6. 

Vaccinia virus is a large double-stranded DNA virus that infects many types of mammalian cells and some invertebrate cells. Its replication cycle is confined to the cytoplasm and can be divided into an early phase and a late phase. The late phase is characterized by the synthesis of structural proteins and enzymes that are packaged into progeny virions. Expression of foreign genes from recombinant vaccinia viruses takes advantage of promoters that are active during the late phase in order to maximize yields of the protein of interest. Several features make the vaccinia virus system... 

Clinical studies with a recombinant vaccine (using Vaccinia virus expressing HPV 16, 18, E6, and E7 proteins) in patients with preinvasive and invasive cancer have been reviewed (1). 

There have already been accusations that two patients with AIDS, treated with an experimental vaccine prepared using a Vaccinia virus that had apparently been inactivated and genetically engineered to express HIV proteins, may have died from vaccinia gangrenosa (14,15). On the other hand, there is evidence that recombinant Vaccinia viruses have reduced pathogenicity (SEDA-13, 289). 

Proteins expressed in bacteria lack post-translational modifications such as phosphorylations and glycosylations. If these are crucial for function, expression in eukaryotic cells is a better choice. Procedures for expression of recombinant proteins in baculovirus or vaccinia virus can be found elsewhere.1516... 

Leffers, H. (1994). Expression of recombinant proteins using vaccinia virus vectors. In Cell Biology A Laboratory Handbook, Vol. 3 (Celis, J.E., ed.), Academic Press, San Diego, pp. 155-163. 

Expression of Recombinant Proteins in the Vaccinia Virus Expression System... 

Stunnenberg, H. G., Lange, H., Philipson, L., van Miltenburg, R. T., and van der Vliet, P. C. (1988) High expression of functional adenovirus DNA polymerase and precursor terminal protein using recombinant vaccinia virus. Nucleic Acids Res. 16, 2431-2444. 

---