Studying the cell biology of apicomplexan parasites using fluorescent proteins

The ability to transfect Apicomplexan parasites has revolutionized the study of this important group of pathogens. The function of specific genes can be explored by disruption of the locus or more subtly by introduction of altered or tagged versions. Using the transgenic reporter gene green fluorescent protein (GFP), cell biological processes can now be studied in living parasites and in real time. We review recent advances made using GFP-based experiments in the understanding of protein trafficking, organelle biogenesis, and cell division in Toxoplasma gondii and Plasmodium falciparum. A technical section provides a collection of basic experimental protocols for fluorescent protein expression in T gondii. The combination of the in vivo marker GFP with an increasingly diverse genetic toolbox for T. gondii opens many exciting experimental opportunities, and emerging applications of GFP in genetic and pharmacological screens are discussed.