Recombinant Human SARS coronavirus Nucleoprotein is produced in E. coli and comprises the full-length sequence of 422 amino acids. This protein carries an N-terminal 6xHis tag for affinity purification, reaching a purity level greater than 90%, as determined by SDS-PAGE. It's intended for research use only and contains no detectable endotoxins, which appears to make it suitable for various experimental applications.
The SARS coronavirus Nucleoprotein (N) seems to play a crucial role in the viral life cycle, particularly in the packaging and assembly of the viral genome. It's involved in viral RNA replication and transcription, making it a key target for research related to viral pathogenesis and immune response. Understanding this protein may be essential for grasping coronavirus biology and developing therapeutic strategies.
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. Antibody Development and Validation
This full-length recombinant SARS-CoV nucleoprotein can serve as an immunogen for generating monoclonal or polyclonal antibodies specific to the viral nucleoprotein. The N-terminal 6xHis tag helps with purification and immobilization for antibody screening assays. ELISA-based screening with this protein allows researchers to identify high-affinity antibodies against different epitopes of the nucleoprotein. The high purity (>90%) likely ensures minimal cross-reactivity during antibody development processes.
2. Protein-Protein Interaction Studies
The nucleoprotein plays crucial roles in viral RNA binding and packaging. This makes the recombinant protein valuable for studying its interactions with viral RNA, other viral proteins, or host cell factors. Pull-down assays can be performed using the 6xHis tag to capture the nucleoprotein and identify binding partners through mass spectrometry or Western blot analysis. Surface plasmon resonance or other biophysical techniques may help quantify binding kinetics and affinities with potential interaction partners.
3. Structural and Biochemical Characterization
This full-length nucleoprotein (1-422aa) expressed in E.coli provides material for detailed structural studies. These might include X-ray crystallography, NMR spectroscopy, or cryo-electron microscopy. The protein can be used to investigate conformational changes, domain organization, and oligomerization states under different buffer conditions. Biochemical assays could examine the protein's stability, folding properties, and response to various environmental conditions relevant to viral replication.
4. Immunoassay Development and Optimization
The recombinant nucleoprotein serves as a reference standard for developing and validating various immunoassays in research settings. The 6xHis tag enables consistent immobilization on nickel-coated surfaces for ELISA development or biosensor applications. This protein appears useful for establishing detection limits, optimizing assay conditions, and validating specificity of newly developed immunoassays for SARS-CoV research.
5. Viral Assembly and RNA Binding Studies
Given the nucleoprotein's role in viral nucleocapsid formation, this recombinant protein can be used in in vitro assembly studies to understand the mechanisms of viral particle formation. RNA-binding specificity and affinity can be investigated using gel shift assays or fluorescence-based binding studies. The protein may also prove useful for studying the kinetics of nucleocapsid assembly and the effects of mutations or small molecule inhibitors on these processes.
