- Description UvsY is the phage T4 recombination mediator protein, and structural and biophysical studies provide insights into its role in T4 homologous recombination. During T4 homologous recombination, the UvsX recombinase must compete with the prebound gp32 single-stranded binding protein for DNA-binding sites and T4 UvsY Protein stimulates this filament nucleation event (1). UvsY is a 15.8-kDa protein with properties that are consistent with its role as mediator; it stimulates the DNA-dependent ATPase activity of UvsX, lowers the critical concentration of UvsX that is required for activity, and promotes strand exchange (2, 3). UvsY efficiently promotes the UvsX-catalyzed strand invasion reaction by recruiting to ssDNA–gp32 complexes, promotes the release of gp32, and favors the binding of UvsX. 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 E. coli BL21 (DE3) strain expressing T4 UvsY gene with an Nterminal His tag. This GST tag does not have any effect on UvsY activity. Product Includes Glycerol Free T4 UvsY protein 10X UvsY Reaction Buffer 1x UvsY reaction buffer composition 20 mM Tris-acetate 100 mM Potassium acetate 10 mM Magnesium acetate 1 mM DTT pH 7.8 @ 25ºC Storage Buffer 50 mM Tris-HCl 50 mM KCl 1 mM DTT 0.1 mM EDTA pH 7.5 @ 25ºC Storage Temperature -20ºC Quality Control assays Glycerol Free T4 UvsY is free from detectable nuclease activities. References 1. Beernink HT, Morrical SW. RMPs: Recombination/replication mediator proteins. Trends Biochem Sci. 1999;24(10):385–389. 2. Hashimoto K, Yonesaki T. The characterization of a complex of three bacteriophage T4 recombination proteins, UvsX protein, UvsY protein, and gene 32 protein, on single-stranded DNA. JBiol Chem. 1991;266(8):4883–4888. 3. Beernink HT, Morrical SW. The UvsY recombination protein of bacteriophage T4 forms hexamers in the presence and absence of single-stranded DNA. Biochemistry. 1998;37(16):5673– 6813572G 3575G 3577G3572GF 3575GF 3577GF
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