Fractionation of Plasmodium chabaudi-infected erythrocytes into parasites and ghosts.
We have developed a new procedure for subfractionation of Plasmodium chabaudi-infected erythrocytes into parasites and ghosts. Trophozoite-infected erythrocytes enriched over a percoll-step (:1.10 g/cm3) are subjected to a glycerol-enhanced osmotic shock. This induces the release of parasites and the emergence of erythrocyte ghosts, which can be separated on a continuous percoll gradient (:1.02----1.10 g/cm3). The parasites are intact in terms of ultrastructure and incorporation of 14C-isoleucine. The erythrocyte ghosts are purified over a two-step percoll gradient (:1.01-1.02 g/cm3). Our method recovers about 40%-50% of the initial ghosts and 70%-95% of the initially freed parasites.
Wunderlich
F
F
Schillinger
G
G
Helwig
M
M
D000818:Animals; D002250:Carbon Radioisotopes; D002458:Cell Fractionation; D004910:Erythrocyte Membrane; D004912:Erythrocytes / Q000469:parasitology*; D051379:Mice; D008854:Microscopy, Electron; D008857:Microscopy, Interference; D009997:Osmotic Pressure; D010274:Parasitology / Q000379:methods*; D010961:Plasmodium / Q000302:isolation & purification*; D011205:Povidone; D012822:Silicon Dioxide
D002250:Carbon Radioisotopes; C016039:Percoll; D012822:Silicon Dioxide; D011205:Povidone