Comparison of the absolute and relative efficiencies of electroporation-based transfection protocols for Plasmodium falciparum.
BACKGROUND
Several electroporation protocols exist to transfect exogenous DNA into Plasmodium falciparum. To date, however, only a subjective analysis of their relative efficiencies has been reported.


METHODS
A time-course of luciferase reporter expression is used to provide an objective quantitative analysis of the absolute efficiency of three electroporation techniques; direct electroporation of ring stage infected erythrocytes, preloading of erythrocytes and a novel "double-tap" protocol that combines both approaches.


RESULTS
Preloading of erythrocytes shows a mean efficiency of 9.59 x 10(-6), some 5-180 fold more efficient than matched experiments utilizing the "double-tap" and direct electroporation of ring stage infected erythrocytes alone, respectively.


CONCLUSIONS
Evidence presented here provides the first quantitative assessment of both the absolute and relative efficiencies of a key molecular tool used to study the biology and pathogenesis of this important human pathogen.
Institute for Science and Technology in Medicine, Keele University, Staffordshire, ST5 5BG, UK.
Sandra
S
Hasenkamp
Karen T
KT
Russell
Paul
P
Horrocks
D018274:Electroporation / Q000379:methods*; D017930:Genes, Reporter; D008156:Luciferases / Q000032:analysis / Q000235:genetics; D008967:Molecular Biology / Q000379:methods*; D010274:Parasitology / Q000379:methods*; D010963:Plasmodium falciparum / Q000235:genetics*; D014162:Transfection / Q000379:methods*
D008156:Luciferases