Analysis of small GTPase function in trypanosomes

Methods Enzymol. 2008;438:57-76. doi: 10.1016/S0076-6879(07)38005-1.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Cloning, Organism / methods
  • Endocytosis / physiology
  • Exocytosis / physiology
  • Genetic Vectors / physiology
  • Molecular Sequence Data
  • Monomeric GTP-Binding Proteins / genetics
  • Monomeric GTP-Binding Proteins / physiology*
  • Protein Transport / physiology
  • RNA Interference
  • Sequence Alignment
  • Transfection / methods
  • Trypanosoma brucei brucei / enzymology*
  • Trypanosoma brucei brucei / growth & development

Substances

  • Monomeric GTP-Binding Proteins