Selection by flow-sorting of genetically transformed, GFP-expressing blood stages of the rodent malaria parasite, Plasmodium berghei.

摘要: This protocol describes a methodology for the genetic transformation of rodent malaria parasite Plasmodium berghei and subsequent selection transformed parasites expressing green fluorescent protein (GFP) by flow-sorting. It provides methods for: transfection schizont stage with DNA constructs that contain gfp as selectable marker; mutants flow-sorting; injection flow-sorted, GFP-expressing into mice collection parasites. The use two different promoters expression GFP is described; these require slightly procedures mutants. enables within 10–12 days after transfection. modification P. widely used to investigate gene function in sp. application flow-sorting increases possibilities mutagenesis, effectively expanding range markers.