Homologous Recombination and Stable Transfection in the Parasitic Protozoan Trypanosoma brucei
- 14 December 1990
- journal article
- Published by American Association for the Advancement of Science (AAAS) in Science
- Vol. 250 (4987), 1583-1587
- https://doi.org/10.1126/science.2177225
Abstract
Development of methods for the manipulation of the genomes of parasitic protozoa will lead to enhanced understanding of parasite biology and host-parasite relationships. Efficient gene transfer and targeted integration by homologous recombination were achieved in the parasitic protozoan Trypanosoma brucei, the causative agent of sleeping sickness. An expression vector with the neomycin phosphotransferase gene (neo), under the control of a procyclic acidic repetitive protein (PARP) gene promoter, was targeted into an intergenic region in beta alpha-tubulin-gene tandem array. Sixteen copies of neo were found in a tandem array in one of the transfectants where the PARP promoter controlled alpha-amanitin-resistant transcription of neo, whereas transcription of tubulin genes remained alpha-amanitin-sensitive.Keywords
This publication has 25 references indexed in Scilit:
- Evidence for segmental gene conversion between a cognate hsp70 gene and the temperature-sensitively transcribed hsp70 genes of Trypanosoma bruceiMolecular and Biochemical Parasitology, 1990
- Stable expression of the bacterial neo r gene in Leishmania enriettiiNature, 1990
- Primer-Directed Enzymatic Amplification of DNA with a Thermostable DNA PolymeraseScience, 1988
- The anatomy and transcription of a telomeric expression site for variant-specific surface antigens in T. bruceiCell, 1987
- DISCONTINUOUS TRANSCRIPTION AND ANTIGENIC VARIATION IN TRYPANOSOMESAnnual Review of Biochemistry, 1986
- The double-strand-break repair model for recombinationCell, 1983
- Tubulin genes are tandemly linked and clustered in the genome of trypanosoma bruceiCell, 1983
- Cloning in single-stranded bacteriophage as an aid to rapid DNA sequencingJournal of Molecular Biology, 1980
- Detection of specific sequences among DNA fragments separated by gel electrophoresisJournal of Molecular Biology, 1975
- Changes in size and secondary structure of the ribosomal transcription unit during vertebrate evolutionJournal of Molecular Biology, 1975