Recombinant Enterobacteria phage T4 Recombination protein uvsY is produced in E. coli with an N-terminal MBP-tag and a C-terminal 6xHis-Avi-tag, which makes detection and purification more straightforward. The protein expresses as a full-length version spanning amino acids 1-137. SDS-PAGE analysis confirms purity levels greater than 85%. This biotinylated protein appears suitable for various research applications, though it's designated for research use only.
The uvsY protein from Enterobacteria phage T4 acts as a recombination mediator protein. It seems to play a crucial role in homologous recombination by helping pair homologous DNA strands. During presynaptic filament assembly, it works alongside other recombination proteins. Scientists studying DNA repair mechanisms and the molecular processes behind genetic exchange in phage biology may find this protein particularly valuable.
Potential Applications
Note: The applications listed below are based on what we know about this protein's biological functions, published research, and experience from experts in the field. However, we haven't fully tested all of these applications ourselves yet. We'd recommend running some preliminary tests first to make sure they work for your specific research goals.
1. Protein-Protein Interaction Studies via Biotin-Streptavidin Capture
Researchers can immobilize the biotinylated uvsY protein on streptavidin-coated surfaces or beads to examine how it interacts with other bacteriophage T4 recombination proteins or host bacterial proteins. The biotin-streptavidin bond is remarkably strong, providing stable protein capture while likely preserving uvsY's native structure for binding studies. This method could help identify and characterize binding partners in T4 recombination pathways through pull-down assays, followed by mass spectrometry or Western blot analysis.
2. ELISA-Based Binding Assays
The biotinylated uvsY may work well as a capture reagent in enzyme-linked immunosorbent assays to measure binding interactions with potential protein partners or DNA substrates. While the biotin tag allows for straightforward immobilization on streptavidin-coated ELISA plates, the MBP and His tags offer additional detection possibilities. This application proves especially useful when screening libraries of potential interacting molecules or conducting dose-response studies to determine binding affinities.
3. Surface Plasmon Resonance (SPR) Analysis
Scientists can immobilize the biotinylated uvsY protein on streptavidin-functionalized SPR sensor chips for real-time kinetic analysis of molecular interactions. This approach allows determination of association and dissociation rate constants, plus equilibrium binding constants for interactions between uvsY and other recombination machinery components. The biotin-streptavidin immobilization strategy appears to provide oriented and stable protein attachment that works well for quantitative binding studies.
4. Affinity Purification of uvsY-Interacting Complexes
This protein's dual-tagged design, featuring both biotin and 6xHis tags, opens up sequential or alternative purification strategies for isolating protein complexes containing uvsY. Researchers might use either streptavidin-based or nickel-based affinity chromatography to purify native complexes from bacteriophage T4-infected bacterial lysates. This strategy could help identify and characterize multi-protein assemblies involved in T4 DNA recombination and repair processes.
5. Fluorescence-Based Binding Studies
The biotinylated uvsY can be linked with fluorescently-labeled streptavidin to create a fluorescent probe for studying protein localization and dynamics in reconstituted recombination systems. This application lets researchers monitor uvsY recruitment to DNA substrates or protein complexes using fluorescence microscopy or fluorescence polarization assays. The approach may prove valuable for understanding how recombination machinery components organize themselves temporally and spatially during DNA repair processes.
