P. falciparum (lab strain DD2) treated with A23187 vs untreated [CGH]

Calcium is a universal second messenger molecule which plays a significant role in several biological processes. Presence of calcium sensors (calmodulins) and calcium-dependent protein kinases in Plasmodium species suggests an important role of calcium-dependent signaling pathways in the regulation of cellular processes in the malaria parasites. Evidence for the transcriptional response of Plasmodium falciparum asexual blood stages to the well-known calcium ionophore A23187 has been presented here. Comparative genomic hybridization (CGH) was carried out with total DNA isolated from the untreated parasites and the parasites treated with the calcium ionophore (A23187 5 μM concentration) at 1 hour, 4 hours and 6 hours post treatment. Genomic DNA from the isolates was extracted by phenol chloroform. 3 μg of the total DNA from each of the isolate was subjected to klenow(NEB) reaction as described in Bozdech Z, Llinas M, Pulliam BL, Wong ED, Zhu J, DeRisi JL: The transcriptome of the intraerythrocytic developmental cycle of Plasmodium falciparum. PLoS Biol 2003, 1(1):E5. Treated DNA labeled with Cy3 was then hybridized against untreated DNA labeled with Cy5.