Characterization of promoters and stable transfection by homologous and nonhomologous recombination in Plasmodium falciparum
- Author(s)
- Crabb, BS; Cowman, AF;
- Details
- Publication Year 1996-07-09,Volume 93,Issue #14,Page 7289-7294
- Journal Title
- PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
- Publication Type
- Journal Article
- Abstract
- Genetic studies of the protozoan parasite Plasmodium falciparum have been severely limited by the inability to introduce or modify genes. In this paper we describe a system of stable transfection of P. falciparum using a Toxoplasma gondii dihydrofolate reductase-thymidylate synthase gene, modified to confer resistance to pyrimethamine, as a selectable marker. This gene was placed under the transcriptional control of the P. falciparum calmodulin gene flanking sequences, Transfected parasites generally main rained plasmids episomally while under selection; however, parasite clones containing integrated forms of the plasmid were obtained, Integration occurred by both homologous and nonhomologous recombination, In addition to the flanking sequence of the P. falciparum calmodulin gene, the 5' sequences of the P. falciparum and P. chabaudi dihydrofolate reductase-thymidylate synthase genes were also shown to be transcriptionally active in P. falciparum. The minimal 5' sequence that possessed significant transcriptional activity was determined for each gene and short sequences containing important transcriptional control elements were identified. These sequences will provide considerable flexibility in the future construction of plasmid vectors to be used for the expression of foreign genes or for the deletion or modification of P. falciparum genes of interest.
- Publisher
- NATL ACAD SCIENCES
- Keywords
- THYMIDYLATE SYNTHASE GENE; HISTIDINE-RICH PROTEIN; TOXOPLASMA-GONDII; CALMODULIN GENE; MALARIA; RESISTANCE; SEQUENCE; DNA; REARRANGEMENT; PYRIMETHAMINE
- Publisher's Version
- https://doi.org/10.1073/pnas.93.14.7289
- Terms of Use/Rights Notice
- Refer to copyright notice on published article.
Creation Date: 1996-07-09 12:00:00