title: Small-scale in vitro culture and purification of Plasmodium berghei for transfection experiment.
abstract: 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.
authors.affiliation: National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), 113 Thailand Science Park, Pathumthani, Thailand.
authors.forename: Voravuth
authors.initials: V
authors.lastname: Somsak
authors.forename: Somdet
authors.initials: S
authors.lastname: Srichairatanakool
authors.forename: Sumalee
authors.initials: S
authors.lastname: Kamchonwongpaisan
authors.forename: Yongyuth
authors.initials: Y
authors.lastname: Yuthavong
authors.forename: Chairat
authors.initials: C
authors.lastname: Uthaipibull
mesh_terms: D000818:Animals; D001769:Blood / Q000469:parasitology; D018929:Cell Culture Techniques; D051379:Mice; D010274:Parasitology / Q000379:methods*; D010962:Plasmodium berghei / Q000235:genetics / Q000254:growth & development*; D014162:Transfection