Site-specific recombination in mammalian cells catalyzed by γδ resolvase mutants: implications for the topology of episomal DNA
| Autor: | Micha Schwikardi, Peter Dröge |
|---|---|
| Jazyk: | angličtina |
| Předmět: |
Tn3 transposon
FLP-FRT recombination Mutant Biophysics Transposases CHO Cells Regulatory Sequences Nucleic Acid Transfection Biochemistry Catalysis Episomal DNA Recombinases Viral Proteins chemistry.chemical_compound Genes Reporter Structural Biology Cricetinae Escherichia coli Genetics Animals γδ resolvase Site-specific recombination Mutant recombinase Molecular Biology Recombination Genetic Integrases biology DNA Superhelical Eukaryote Cell Biology biology.organism_classification Molecular biology DNA topology chemistry Attachment Sites Microbiological Mutation Nucleic Acid Conformation DNA supercoil Recombination DNA Plasmids |
| Zdroj: | FEBS Letters. (2-3):147-150 |
| ISSN: | 0014-5793 |
| DOI: | 10.1016/S0014-5793(00)01394-6 |
| Popis: | We have transferred the prokaryotic gammadelta resolvase system to mammalian cells and present a comparative analysis of recombination by wild-type and two mutant resolvases (E124Q and E102Y/E124Q). Transient co-transfection assays using beta-galactosidase as reporter for recombination reveal that episomal DNA does not contain a significant level of unconstrained negative supercoiling, since only mutant resolvases are recombination-proficient. We also show that the efficiency of recombination by the resolvase double mutant is comparable to that observed with Cre, which indicates that resolvase can be used as a new tool for controlled manipulations of episomal DNAs. |
| Databáze: | OpenAIRE |
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