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Separation of DNA‐containing organelles from Toxoplasma gondii by CZE
Author(s) -
Moe Morten K.,
Samuelsen PerJostein,
Nielsen Henrik V.,
Nielsen Kaare M.
Publication year - 2010
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200900582
Subject(s) - apicoplast , toxoplasma gondii , organelle , apicomplexa , lysis , biology , mitochondrial dna , dna , mitochondrion , nucleic acid , microbiology and biotechnology , biochemistry , plasmodium falciparum , genetics , gene , malaria , antibody , immunology
Toxoplasma gondii and other members of the family Apicomplexa have two organelles, in addition to the nucleus, that contain DNA. Herein is reported the separation of the DNA‐carrying organelles from T. gondii tachyzoites, i.e. the mitochondrion and the apicoplast, by CZE. The cells were stained with SYTO9, a dye that exhibit fluorescence when interacting with double stranded nucleic acids ( e.g. DNA) and disrupted by nitrogen cavitation. Following careful removal of the heavier cellular material, the remaining lysate was injected on a CE instrument and the DNA‐containing organelles were detected by LIF. The mitochondrion had longer migration time than the apicoplast, and the migration times were comparable in the replicates. This method should potentially also work for other members of the Apicomplexa including Plasmodium falciparum .