Recombinant Bovine coronavirus Non-structural protein 2a (2a) is produced using an E. coli expression system and spans the complete protein sequence from amino acids 1 to 278. The protein includes a C-terminal 6xHis-tag that makes purification more straightforward. SDS-PAGE analysis shows the protein reaches greater than 85% purity, which appears suitable for most research applications. This product is strictly for research use and cannot be used for therapeutic or diagnostic purposes.
Non-structural protein 2a seems to play a role in how Bovine coronavirus replicates and transcribes its genetic material. As part of the virus's replication machinery, it likely serves an important function in the viral life cycle. Studying this protein may be critical for virology research and could potentially help in developing antiviral strategies. It has become a focus for researchers investigating how coronaviruses replicate and cause disease.
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.
While full-length expression preserves the primary sequence, E. coli often fails to correctly fold complex viral NSPs—many of which require precise disulfide bonds, transmembrane interactions, or host factor binding for function. No data confirms native secondary/tertiary structure (e.g., circular dichroism for disulfide bonds, thermal shift assays for stability). E. coli may produce misfolded NSP2a, leading to aggregation or non-functional conformations. NSP2a is critical for viral replication (e.g., forming the replication-transcription complex with other NSPs). Its bioactivity (e.g., binding to NSP12/NSP7 or host factors) is untested—E. coli-expressed protein may lack functional interfaces, limiting its ability to mimic native NSP2a.
1. Antibody Development and Immunoassay Research
This recombinant NSP2a can serve as an immunogen for generating antibodies, but antibody specificity must be validated against native NSP2a—E. coli-expressed protein may present non-native epitopes, leading to cross-reactivity with irrelevant targets. The His tag simplifies purification/immobilization for ELISA, but antibodies may not recognize the fragment in its native context (e.g., in infected cells). High purity supports consistent immunogenicity, but validation with native protein is critical.
2. Protein-Protein Interaction Studies
Pull-down assays using the His tag can identify interactors, but results depend on correct folding—E. coli-expressed NSP2a may misfold, causing false positives/negatives. Identified partners (e.g., other NSPs or host factors) must be validated via co-IP or functional assays (e.g., replicon assays) to rule out artifacts. The full length preserves interaction domains, but bioactivity (e.g., binding to NSP3) is unconfirmed.
3. Biochemical Characterization and Structural Studies
This NSP2a supports preliminary biophysical studies (e.g., molecular weight confirmation, thermal stability assays, circular dichroism for secondary structure) but cannot fully represent native NSP2a—E. coli-expressed protein may lack correct disulfide bonds or oligomerization states. Structural conclusions must contextualize folding limitations, and the His tag may interfere with high-resolution techniques (e.g., crystallography).
4. Serological Research Tools
This recombinant NSP2a can function as an antigen for serological assays, but its native conformation must be confirmed—E. coli-expressed protein may misfold, altering epitope presentation. ELISA protocols using this antigen may yield inaccurate results if the protein does not mimic native NSP2a. Validate antigenicity with native NSP2a or infected animal sera before use in epidemiological studies.
Final Recommendation & Action Plan
This E. coli-expressed BCoV NSP2a has potential for antibody development or preliminary biochemical studies, but requires rigorous validation first, confirm folding via circular dichroism/thermal shift assays to rule out misfolding; second, validate bioactivity (e.g., binding to NSP12) using co-IP or replicon assays; third, for serological tools, confirm antigenicity with native protein. Optimize expression (e.g., co-express chaperones like GroEL/ES) to improve native folding. If folding/bioactivity fails, switch to a eukaryotic system (e.g., insect cells) to ensure native structure—this NSP2a’s utility for functional studies depends on validating its structural and functional integrity.
