Conditional mutagenesis using site-specific recombination in Plasmodium berghei.
Reverse genetics in Plasmodium, the genus of parasites that cause malaria, still faces major limitations. Only red blood cell stages of this haploid parasite can be transfected. Consequently, the function of many essential genes in these and subsequent stages, including those encoding vaccine candidates, cannot be addressed genetically. Here, we establish conditional mutagenesis in Plasmodium by using site-specific recombination and the Flp/FRT system of yeast. Site-specific recombination is induced after cross-fertilization in the mosquito vector of two clones containing either the target sequence flanked by two FRT sites or the Flp recombinase. Parasites that have undergone recombination are recognized in the cross progeny through the expression of a fluorescence marker. This approach should permit to dissect the function of any essential gene of Plasmodium during the haploid phase of its life, i.e., during infection of salivary glands in the mosquito and infection of both the liver and red blood cells in the mammal.
Carvalho
Teresa Gil
TG
Unité de Biologie et Génétique du Paludisme, Institut Pasteur, 25 Rue du Docteur Roux, 75724 Paris Cedex 15, France.
Thiberge
Sabine
S
Sakamoto
Hiroshi
H
Ménard
Robert
R
D000818:Animals; D003433:Crosses, Genetic; D005306:Fertilization; D005838:Genotype; D006801:Humans; D016296:Mutagenesis; D010957:Plasmids / Q000235:genetics; D010962:Plasmodium berghei / Q000235:genetics* / Q000502:physiology; D011995:Recombination, Genetic; D017384:Sequence Deletion; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics; D014162:Transfection
D013762:Tetrahydrofolate Dehydrogenase