Analysis of small GTPase function in trypanosomes.
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.
Field
Mark C
MC
Department of Pathology, University of Cambridge, Cambridge, United Kingdom.
Horn
David
D
Carrington
Mark
M
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
D020559:Monomeric GTP-Binding Proteins