27] Vaccinia virus as vector to express ion channel genes

Abstract

This chapter describes the use of vaccinia virus (VV)—a lytic DNA virus—to achieve the transient expression of voltage-gated ion channels in various primary cell cultures and cell lines. Recombinant VV vectors present several advantages for eukaryotic expression technology, such as VV particles have a complex morphology with an outer envelope surrounding the biconcave core and two lateral bodies of unknown function and the genome consists of a linear, double- stranded DNA molecule of 187 kilobases (kb), packaged within the virus core. Another advantage is that VV is able to infect 100% of a eukaryotic cell population. The principal advantage of recombinant VV over other transient expression systems is the broad range of mammalian cell types that can be infected at high success rates. Thus, VV may prove extremely useful in the expression of ion channel and receptor/ionophore proteins that require cell-specific posttranslational processing, or when cell-specific subunits of these excitability proteins are required for complete functional reconstitution.