This recombinant DNA-binding protein HU-beta from Salmonella typhi is expressed in E.coli and includes the full-length sequence from 1-90 amino acids. The protein carries an N-terminal 10xHis-SUMO tag and a C-terminal Myc tag, which streamlines purification and detection steps. Purity appears to exceed 90% as verified by SDS-PAGE, providing what seems to be a reliable reagent for research applications. This product is intended for research use only.
HU-beta is a DNA-binding protein that may play a crucial role in organizing and compacting bacterial chromosomal DNA. It's known for its ability to bend DNA, which likely influences processes such as replication, transcription, and recombination. As one of the nucleoid-associated proteins, HU-beta could be significant in studies examining DNA architecture and gene regulation in prokaryotic systems.
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.
Salmonella typhi DNA-binding protein HU-beta is a bacterial nucleoid-associated protein that requires precise folding, proper dimerization, and specific tertiary structure for its functional activity in DNA binding and nucleoid organization. The E. coli expression system is homologous to this bacterial protein, which increases the probability of correct folding. However, the large N-terminal 10xHis-SUMO tag (∼15 kDa) and C-terminal Myc-tag are significantly larger than the protein itself (90 aa, ∼10 kDa), creating severe steric interference that may disrupt dimerization and DNA-binding domains. While the full-length protein (1-90aa) contains all functional domains, the probability of correct folding with functional DNA-binding activity is low without experimental validation of dimerization and DNA-binding capability.
1. DNA-Protein Interaction Studies Using Electrophoretic Mobility Shift Assays (EMSA)
This application carries a significant risk without functional validation. HU-beta's DNA-binding activity requires precise dimerization and proper tertiary structure. If correctly folded and active (verified through DNA-binding assays), the protein may be suitable for EMSA studies. If misfolded/inactive (unverified), binding data will yield biologically meaningless results due to potential steric interference from the large tags.
2. Antibody Development and Immunoassay Optimization
This application is highly suitable as antibody development relies on antigenic sequence recognition rather than functional protein folding. The full-length protein provides comprehensive epitope coverage for generating HU-beta-specific antibodies. The high purity (>90%) ensures minimal contamination-related issues during immunization protocols.
3. Protein-Protein Interaction Mapping via Pull-Down Assays
This application requires proper folding and dimerization validation. HU-beta interactions with other nucleoid proteins require native conformation. If correctly folded (verified), the protein may identify physiological interaction partners. If misfolded/unverified, there is a high risk of non-specific binding or tag-mediated artefacts.
4. Comparative Structural and Functional Analysis
Meaningful comparative studies require native protein conformation and functional activity. If correctly folded and active (verified), the protein enables valid evolutionary comparisons. If misfolded/inactive (unverified), comparative analyses would yield misleading insights about HU protein conservation and function.
Final Recommendation & Action Plan
The E. coli-expressed HU-beta with large dual tags poses challenges for functional studies due to the severe steric interference from tags relative to the small protein size. Begin with biophysical characterization (e.g., size-exclusion chromatography for dimerization analysis, circular dichroism spectroscopy for secondary structure) and validate DNA-binding activity using EMSA before considering functional applications. Applications 2 (antibody development) can proceed immediately. Applications 1, 3, and 4 require rigorous functional validation. For reliable HU-beta research, consider tag removal or use tag-free constructs to minimize interference with dimerization and DNA-binding domains.
