Homologous recombination is important for the error-free repair of DNA double-strand breaks and for replication fork restart. Recombinases of the RecA/RAD51 family perform the central catalytic role in this process. UvsX recombinase is the RecA/Rad51 ortholog of bacteriophage T4. T4 UvsX DNA Recombinase and other recombinases form presynaptic filaments on ssDNA that are activated to search for homology in dsDNA and to perform DNA strand exchange (1-3).
Note:Glycerol acts as a cryoprotectant and protein stabilizer when added to the storage buffer of proteins and enzymes. However, in certain circumstances, it is preferred to omit glycerol from the buffer. This includes instances where the presence of glycerol may interfere, such as lyophilization, high-throughput instruments with sensitive fluidics or primary cell cultures.For our glycerol free products, it is recommended to use immediately upon thaw as without glycerol there will be significant activity loss with freeze/thaw cycles.
Protein Purity
The physical purity of this enzyme is ≥98% as assessed by SDS-PAGE with Coomassie® blue staining (Fig. 1).
Product Source
- Ecoli BL21 (DE3) strain expressing T4 UvsX gene.
Product Includes
- Glycerol Free T4 UvsX Recombinase
- 10X UvsX Recombinase Reaction Buffer
1x UvsX Recombinase reaction buffer composition
20 mM Tris-acetate pH 7.8100 mM Potassium acetate10 mM Magnesium acetate1 mM DTT
Storage Buffer
50 mM Tris-HCl50 mM KCl1 mM DTT0.1 mM EDTApH 7.5 @ 25ºC
Storage Temperature
–20ºC
Quality Control assays
Glycerol Free T4 UvsX DNA Recombinase is free from detectable nuclease activities.
References
- Cromie GA, Connelly JC, Leach DR (2001) Recombination at double-strand breaks and DNA ends: conserved mechanisms from phage to humans. Mol Cell 8: 1163–1174
- Michel B, Grompone G, Flores MJ, Bidnenko V (2004) Multiple pathways process stalled replication forks. Proc Natl Acad Sci U S A 101: 12783–12788
- Liu J, Ehmsen KT, Heyer WD, Morrical SW (2011) Presynaptic filament dynamics in homologous recombination and DNA repair. Crit Rev Biochem Mol Biol 46: 240–270
3562G 3565G
