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Integration of a transfected gene into the genome of Babesia bovis occurs by legitimate homologous recombination mechanisms
- Source :
- Molecular and Biochemical Parasitology. 202:23-28
- Publication Year :
- 2015
- Publisher :
- Elsevier BV, 2015.
-
Abstract
- This study examines the patterns of gene integration of gfp-bsd upon stable transfection into the T3Bo strain of Babesia bovis using a plasmid designed to integrate homologous sequences of the parasite's two identical ef-1α A and B genes. While the transfected BboTf-149-6 cell line displayed two distinct patterns of gene integration, clonal lines derived from this strain by cell sorting contained only single gfp-bsd insertions. Whole genome sequencing of two selected clonal lines, E9 and C6, indicated two distinct patterns of gfp-bsd insertion occurring by legitimate homologous recombination mechanisms: one into the expected ef-1α orf B, and another into the ef-1α B promoter. The data suggest that expression of the ef-1α orf B is not required for development of B. bovis in cultured erythrocyte stages. Use of legitimate homologous recombination mechanisms in transfected B. bovis supports the future use of transfection methods for developing efficient gene function assignment experiments using gene knockout techniques.
- Subjects :
- Erythrocytes
Genes, Protozoan
Green Fluorescent Proteins
Cattle Diseases
Biology
Transfection
Genome
Gene Knockout Techniques
Peptide Elongation Factor 1
Plasmid
Babesiosis
Animals
Homologous Recombination
Promoter Regions, Genetic
Molecular Biology
Gene
Whole genome sequencing
Genetics
Babesia bovis
Sequence Analysis, DNA
biology.organism_classification
Molecular biology
Clone Cells
Mutagenesis, Insertional
Cattle
Parasitology
Homologous recombination
Subjects
Details
- ISSN :
- 01666851
- Volume :
- 202
- Database :
- OpenAIRE
- Journal :
- Molecular and Biochemical Parasitology
- Accession number :
- edsair.doi.dedup.....ad31e5b872798858d2c8e696a69490c3