Experimental Approaches for Examining Apicoplast Biology
Genetic manipulation is a powerful tool to study gene function but identifying the direct and primary functional outcomes of any gene depletion is crucial for this strategy to be productive. This is a major challenge for the study of apicoplast biology, because, in the absence of an efficient isolation method, apicoplast functions must be assayed in the parasite. These assays should be performed dynamically from the time of gene depletion, and include standards and controls that separate direct from indirect phenotypes. Here, we describe a pipeline for apicoplast functional analysis and highlight relevant mutant T. gondii cell lines and apicoplast markers that are available in the field and that enhance the specificity of phenotype description.
Key wordsApicoplast Plastid Import Organelle Redox Fluorescence roGFP qPCR qRT-PCR Live imaging
- 4.Biddau M, Bouchut A, Major J, Saveria T, Tottey J, Oka O, van-Lith M, Jennings KE, Ovciarikova J, DeRocher A, Striepen B, Waller RF, Parsons M, Sheiner L (2018) Two essential Thioredoxins mediate apicoplast biogenesis, protein import, and gene expression in Toxoplasma gondii. PLoS Pathog 14(2):e1006836CrossRefGoogle Scholar