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  • 1
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    Stable transfection of malaria parasite blood stages (1995)
    American Association for the Advancement of Science (AAAS)
    Details
    Publication Date: 1995-06-02
    Description: Genetic manipulation of malaria parasites would revolutionize the study of this group of pathogens and have implications for vaccine and drug development. This report describes the stable, drug-selectable genetic transformation of the clinically relevant intracellular blood stages of a malaria parasite. A plasmid transfection vector carrying the gene locus that encodes a drug-resistant form of the bifunctional enzyme dihydrofolate reductase-thymidylate synthase from the rodent malaria parasite Plasmodium berghei was constructed. Derivatives of this vector were introduced into merozoites of P. berghei by electroporation, and parasites were selected for successful transformation in the rodent host on the basis of resistance to pyrimethamine. The plasmids were present in a circular, unrearranged form that replicated episomally to an observed maximum of 15 copies per cell in drug-resistant populations.〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉van Dijk, M R -- Waters, A P -- Janse, C J -- New York, N.Y. -- Science. 1995 Jun 2;268(5215):1358-62.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉Department of Parasitology, University of Leiden, Netherlands.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/7761856" target="_blank"〉PubMed〈/a〉
    Keywords: Animals ; Base Sequence ; DNA Replication ; Drug Resistance ; Electroporation ; Erythrocytes/parasitology ; Genes, Protozoan ; Genetic Vectors ; Molecular Sequence Data ; Multienzyme Complexes/*genetics ; Plasmids ; Plasmodium berghei/drug effects/*genetics/growth & development ; Point Mutation ; Pyrimethamine/*pharmacology ; Rats ; Rats, Wistar ; Replication Origin ; Tetrahydrofolate Dehydrogenase/*genetics ; Thymidylate Synthase/*genetics ; *Transfection
    Print ISSN: 0036-8075
    Electronic ISSN: 1095-9203
    Topics: Biology , Chemistry and Pharmacology , Computer Science , Medicine , Natural Sciences in General , Physics
    Published by American Association for the Advancement of Science (AAAS)
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  • 2
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    Expression of a Plasmodium gene introduced into subtelomeric regions of Plasmodium berghei chromosomes (1996)
    American Association for the Advancement of Science (AAAS)
    Details
    Publication Date: 1996-02-02
    Description: Targeted integration of exogenous DNA into the genome of malaria parasites will allow their phenotype to be modulated by means of gene disruption or the stable expression of foreign and mutated genes. Described here is the site-specific integration through reciprocal exchange, and subsequent expression, of a selectable marker gene into the genome of the pathogenic, bloodstage forms of the rodent malaria parasite Plasmodium berghei. Stable integration of a single copy of the marker gene (retained for more than 70 generations in the absence of drug pressure) into a nontranscribed subtelomeric repeat array of different chromosomes was observed. Expression of the gene within the subtelomeres indicated that the previously recorded absence of transcription in these regions could be due to a corresponding absence of genes rather than active silencing mechanisms.〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉van Dijk, M R -- Janse, C J -- Waters, A P -- New York, N.Y. -- Science. 1996 Feb 2;271(5249):662-5.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉Department of Parasitology, University of Leiden, Netherlands.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/8571132" target="_blank"〉PubMed〈/a〉
    Keywords: Animals ; Antimalarials/pharmacology ; Chromosomes/genetics ; Drug Resistance ; Electroporation ; Gene Expression ; *Genes, Protozoan ; Genetic Vectors ; Multienzyme Complexes/*genetics ; Plasmids ; Plasmodium berghei/drug effects/enzymology/*genetics ; Pyrimethamine/pharmacology ; Repetitive Sequences, Nucleic Acid ; Telomere/*genetics ; Tetrahydrofolate Dehydrogenase/*genetics ; Thymidylate Synthase/*genetics ; *Transfection
    Print ISSN: 0036-8075
    Electronic ISSN: 1095-9203
    Topics: Biology , Chemistry and Pharmacology , Computer Science , Medicine , Natural Sciences in General , Physics
    Published by American Association for the Advancement of Science (AAAS)
    Location Call Number Expected Availability
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    PAPER CURRENT
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