FEBS letters, 2000-04, Vol.471 (2-3), p.147-150
2000
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Details
- Autor(en) / Beteiligte
- Titel
- Site-specific recombination in mammalian cells catalyzed by gammadelta resolvase mutants: implications for the topology of episomal DNA
- Ist Teil von
- FEBS letters, 2000-04, Vol.471 (2-3), p.147-150
- Ort / Verlag
- England
- Erscheinungsjahr
- 2000
- Quelle
- Wiley-Blackwell Journals
- Beschreibungen/Notizen
- 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.
- Sprache
- Englisch
- Identifikatoren
- ISSN: 0014-5793Titel-ID: cdi_proquest_miscellaneous_71035365
- Format
- –
- Schlagworte
- Animals, Attachment Sites, Microbiological - genetics, Catalysis, CHO Cells, Cricetinae, DNA, Superhelical - chemistry, DNA, Superhelical - genetics, DNA, Superhelical - metabolism, Escherichia coli - enzymology, Escherichia coli - genetics, Genes, Reporter - genetics, Integrases - genetics, Integrases - metabolism, Mutation - genetics, Nucleic Acid Conformation, Plasmids - chemistry, Plasmids - genetics, Plasmids - metabolism, Recombinases, Recombination, Genetic - genetics, Regulatory Sequences, Nucleic Acid - genetics, Transfection, Transposases - genetics, Transposases - metabolism, Viral Proteins