Project description:The purpose of this reasearch is to identify and evaluate the global gene expression of Plasmodium knowlesi blood-stage parasites and specifically compare the gene expression profiles of samples derived from ex vivo versus long-term in vitro cultures. For ex vivo samples, a rhesus monkey was infected with P. knowlesi to obtain ring iRBCs to establish a short-term culture to immediately collect ex vivo derived time points every 4 hours in the course of the parasite’s 24-hour life cycle. For in vivo samples, they were generated from the same clone in the ex vivo culture adapted to long-term in vitro culture and were collected every 4 hours in the course of the parasite's 24-hour life cycle. Two replicate experiments(Aa and Ha) were developed from in vitro cultures. Samples labeled with Cy5 were hybridized against a reference RNA pool labeled with Cy3, consisting of equal amounts of P. knowlesi RNA from each time point.
Project description:The purpose of this research is to identify and evaluate the global gene expression of the rodent malaria parasites Plasmodium yoelii, Plasmodium berghei and Plasmodium chabaudi blood-stage parasites and specifically compare the blood stage gene expression profiles of samples derived from previous studies on Plasmodium falciparum, Plasmodium vivax and Plasmodium knowlesi Overall design: Mice were infected by intraperitoneal injections of P. berghei ANKA, P. chabaudi AS or P. yoelii 17x parasitized erythrocytes and parasitaemia and parasite stages were monitored by thin blood smears stained with Giemsa. Mice infected with P. chabaudi were highly synchronized and terminal bled every 2 hours under anesthesia over the course of 24 hr. For P. berghei and P. yoelii infection, mice were terminal bleed and the stage-specific parasitized erythrocytes were separated via Nycodenz density gradient. The ring stage interface was isolated, washed and subjected to ex vivo culture, which was then collected every 2 hr over the course of 24 hr over a complete IDC life-cycle. Samples labeled with Cy5 were hybridized against a reference RNA pool labeled with Cy3, consisting of equal amounts of P. yoelii or P. berghei or P. chabaudi RNA from each time point.