Somsak Voravuth, Srichairatanakool Somdet, Kamchonwongpaisan Sumalee, Yuthavong Yongyuth, Uthaipibull Chairat
National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), 113 Thailand Science Park, Pathumthani, Thailand.
Mol Biochem Parasitol. 2011 Jun;177(2):156-9. doi: 10.1016/j.molbiopara.2011.01.013. Epub 2011 Feb 1.
The standard protocol for genetic modification of the rodent malaria parasite Plasmodium berghei requires infected blood from one or more laboratory mice, followed by large-scale in vitro parasite culture and purification of mature schizonts. Here, protocols are described for small-scale in vitro culture from 20 μL of mouse tail blood and purification of mature P. berghei schizonts sufficient for a single transfection experiment. All procedures are performed in 1.5-mL microcentrifuge tubes. We confirmed that transgenic parasites could be obtained using schizonts prepared by this protocol. This small-scale protocol provides significant advantages, namely reduction of parasite sample, laboratory consumables and mice for transfection experiments.
