Abstract
Leishmania is a genus of trypanosomatid protozoa and is the parasite responsible for the disease leishmaniasis. These protozoa, regulate their gene expression in an atypical way, compared to other higher eukaryotes. The regulation of gene expression is characterized by a predominance of post-transcriptional over pre-transcriptional regulatory mechanisms (Clayton, 2002). Thus proteomic analysis has proven an essential tool for understanding pathways implicated in Leishmania infectivity, host-parasite interactions, drug resistance and others. When employing a comparative proteomics analysis between different parasitic cell lines, it is essential that these lines are cultivated in exactly the same way, in the same cell density and growth phase. More importantly when cell-cycle defects are suspected, it is essential to synchronize cell-lines in the same cell-cycle phase so as to eliminate possible artifacts. This protocol describes the preparation of whole-protein samples for proteomic analysis in Leishmania donovani (L. donovani).
Keywords: Leishmania, Proteomics, Sample preparation
Materials and Reagents
Equipment
Procedure
Notes
Recipes
Acknowledgments
This protocol was adapted from Clayton (2002) and Smirlis et al. (2006). Partial funding from the General Secretariat of Research and Technology action “Proposals for the development of Research Institutes, Krepis” supported this work.
References
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