title: Analysis of small GTPase function in trypanosomes.
abstract: Trypanosomatids are protozoan parasites, of interest due to both their disease burden and deeply divergent position within the eukaryotic lineage. The African trypanosome, Trypanosoma brucei, has emerged as a very amenable model system, with a considerable toolbox of methods available, including inducible overexpression, RNA interference, and a completed genome. Here we describe some of the special considerations that need to be addressed when studying trypanosome gene function, and in particular small GTPases; we provide protocols for transfection, RNA interference, overexpression and basic transport assays, in addition to an overview of available vectors, cell lines, and strategies.
authors.affiliation: Department of Pathology, University of Cambridge, Cambridge, United Kingdom.
authors.forename: Mark C
authors.initials: MC
authors.lastname: Field
authors.forename: David
authors.initials: D
authors.lastname: Horn
authors.forename: Mark
authors.initials: M
authors.lastname: Carrington
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D019976:Cloning, Organism / Q000379:methods; D004705:Endocytosis / Q000502:physiology; D005089:Exocytosis / Q000502:physiology; D005822:Genetic Vectors / Q000502:physiology; D008969:Molecular Sequence Data; D020559:Monomeric GTP-Binding Proteins / Q000235:genetics / Q000502:physiology*; D021381:Protein Transport / Q000502:physiology; D034622:RNA Interference; D016415:Sequence Alignment; D014162:Transfection / Q000379:methods; D014346:Trypanosoma brucei brucei / Q000201:enzymology* / Q000254:growth & development
chemical_list: D020559:Monomeric GTP-Binding Proteins