Green fluorescent protein as a marker in Plasmodium berghei transformation.
We present a new marker that confers both resistance to pyrimethamine and green fluorescent protein-based fluorescence on the malarial parasite Plasmodium berghei. A single copy of the cassette integrated into the genome is sufficient to direct fluorescence in parasites throughout the life cycle, in both its mosquito and vertebrate hosts. Erythrocyte stages of the parasite that express the marker can be sorted from control parasites by flow cytometry. Pyrimethamine pressure is not necessary for maintaining the cassette in transformed parasites during their sporogonic cycle in mosquitoes, including when it is borne by a plasmid. This tool should thus prove useful in molecular studies of P. berghei, both for generating parasite variants and monitoring their behavior.
Sultan
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Michael Heidelberger Division of Immunology, Department of Pathology, Kaplan Cancer Center, New York University Medical Center, New York, New York 10016, USA.
Thathy
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Nussenzweig
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V
Ménard
R
R
D000818:Animals; D000962:Antimalarials / Q000494:pharmacology; D001483:Base Sequence; D002460:Cell Line; D009033:Culicidae / Q000469:parasitology; D017931:DNA Primers / Q000235:genetics; D004351:Drug Resistance / Q000235:genetics; D004912:Erythrocytes / Q000469:parasitology; D005453:Fluorescence; D005819:Genetic Markers; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics*; D010962:Plasmodium berghei / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development; D011739:Pyrimethamine / Q000494:pharmacology; D051381:Rats; D014170:Transformation, Genetic
D000962:Antimalarials; D017931:DNA Primers; D005819:Genetic Markers; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D011739:Pyrimethamine