Study of Nucleolar Localization of Adenovirus Core Proteins

This chapter describes the techniques used to study nucleolar-localized proteins. The chapter starts with cloning of viral proteins for expression in mammalian cells as fusion proteins to well-characterized tags such as enhanced green fluorescence protein (EGFP). This follows on to techniques for transient expression in mammalian cells and immunofluorescence techniques used to examine subcellular localization. Finally there is guidance on the types of antigens and metabolic features of the nucleolus that can be used as markers to confirm that the protein in question is indeed localized in the nucleolus and determine whether it affects gross rRNA synthesis.