4] Construction and use of recombinant vaccinia virus vectors

Abstract

This chapter describes the construction and use of recombinant vaccinia virus (VV) vectors. It presents comments on the specific techniques and methodology required to use this system. New techniques in genetic engineering and molecular biology have provided several novel approaches for the construction and use of recombinant eukaryotic vector systems. These include a variety of mammalian expression plasmids and a number of different animal virus vectors, among which vaccinia virus (VV) is the most well known. VV has been used as a eukaryotic cloning and expression vector for the study of gene systems and protein expression as well as for the development of new vaccines. The development of VV as a vector has been fueled by basic research into the replication of poxviruses, which has identified many attributes contributing to its use as recombinant vectors or vaccines. VV has been used as a live vaccine against smallpox for over 200 years resulting in the eradication of this disease. Vaccinia virus is related to variola virus, the causative agent of smallpox but does not itself cause disease. It is not related to varicella virus, a herpesvirus that causes chickenpox. The expression of products from recombinant VV may be modulated by a number of variables. Specifically, promoters used, mRNA stability, protein stability, multiplicity of infection, time course of the infection, and cell type all affect product yield. VV has been, and continues to be, a powerful tool for research on gene systems and their expression and regulation.