Table 1.

Examples of molecular tools in T. brucei. There are at least eight drugs for selection (G418, Hygromycin, Puromycin, Phleomycin, Blasticidin, Nourseothricin/ClonNAT, Ganciclovir and FOA). Cells are typically grown in semi-defined media (SDM-79 for procyclic form [55], HMI-9 for bloodstream form [56]). Procyclic form cells readily grow up to a density of 1 × 10⁷ cells ml⁻¹ (1 × 10⁶ cells ml⁻¹ for bloodstream form cells) and can be frozen for long-term storage in liquid nitrogen. A subspecies, Trypanosoma brucei brucei, cannot infect humans owing to its sensitivity to human lytic factor [57], and is used in many research laboratories. Various monoclonal antibodies are also available [58]. Genetic exchange occurs under special circumstances (in the tsetse fly [59–61]), but it is not a widely practicable technique. Differentiation of life cycles can be reproduced in vitro [62–64]. GFP, green fluorescent protein; TAP, tandem affinity purification; YFP, yellow fluorescent protein.

techniques	references
epitope-tagging (e.g. TAP, FLAG, GFP and YFP) and gene deletion using homologous recombination	[65–69]
regulated gene expression using TetR and T7 RNA polymerase	[70–72]
Cre-Lox recombination	[73,74]
RNAi, genome-wide RNAi screening	[75–78]
fluorescence in situ hybridization	[79]
GFP tagging of chromosomes using LacO/LacI	[80,81]
affinity purification (immunoprecipitation, BioID)	[82–86]
chromatin immunoprecipitation (ChIP), ChIP-seq	[87,88]
microtubule drugs	[89–91]
live-cell imaging	[92–94]
stable isotope labelling by amino acids in cell culture	[95–97]