Comparison of the absolute and relative efficiencies of electroporation-based transfection protocols for Plasmodium falciparum.

Horrocks

doi:10.1186/1475-2875-11-210

Comparison of the absolute and relative efficiencies of electroporation-based transfection protocols for Plasmodium falciparum.

Horrocks

Authors

Paul Horrocks p.d.horrocks@keele.ac.uk

Abstract

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. CONCLUSION: 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.

Acceptance Date	Jun 21, 2012
Publication Date	Jan 1, 2012
Journal	Malaria Journal
Publisher	Springer Verlag
Pages	210 - ?
DOI	https://doi.org/10.1186/1475-2875-11-210
Keywords	luciferase, electroporation, transfection efficiency, malaria
Publisher URL	http://www.malariajournal.com/content/11/1/210