Analysis of Plasmodium falciparum mitochondrial six-kilobase DNA by pulse-field electrophoresis.
The 6-kb mtDNA of Plasmodium falciparum is thought to replicate by a recombination-dependent mechanism generating large complex branched structures. For technical reasons, including shearing caused by DNA extraction methods, a meaningful quantitative comparison of large complex mtDNA forms has not been feasible. With the use of pulse-field gel electrophoresis, which minimizes any loss or shearing of DNA, we were able to identify an unusually slow migrating population of mtDNA that was resolved from the 6-23-kb population of linear concatemers. Levels of this slow-migrating population of mtDNA were highest during early schizont stage, suggesting that these forms represent replication intermediates. This approach provides a convenient means to monitor the presence of large mtDNA structures in P. falciparum.
Weissig
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Center for Imaging and Pharmaceutical Research, Massachusetts General Hospital, Charlestown 02129, USA.
Rowe
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TC
D000818:Animals; D015139:Blotting, Southern; D004272:DNA, Mitochondrial / Q000032:analysis* / Q000302:isolation & purification; D016054:DNA, Protozoan / Q000032:analysis; D016521:Electrophoresis, Gel, Pulsed-Field / Q000379:methods*; D010963:Plasmodium falciparum / Q000235:genetics*
D004272:DNA, Mitochondrial; D016054:DNA, Protozoan