Recombinant Rickettsia conorii Outer membrane protein B (ompB) is expressed in E. coli, covering the 1363-1655 amino acid region of the protein. This partial protein carries a C-terminal 6xHis-tag, which helps with purification and detection. The product achieves a high purity level of over 90%, as confirmed by SDS-PAGE analysis. This purity appears sufficient for precise research applications.
Outer membrane protein B (ompB) from Rickettsia conorii plays a crucial role in the bacterial outer membrane structure. It seems to be involved in the infection process and represents a key target for studying Rickettsia pathogenesis. Researchers work with ompB to investigate host-pathogen interactions and understand the mechanisms of bacterial adhesion and invasion. This protein is likely integral to advancing research in microbial pathogenesis and infectious diseases.
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.
Based on the provided information, the recombinant Rickettsia conorii ompB fragment is expressed in E. coli, a prokaryotic system that is generally suitable for producing bacterial outer membrane proteins. As both Rickettsia and E. coli are prokaryotes, the protein has a reasonable probability of proper folding. The expressed fragment (1363-1655aa, 292 amino acids) represents a partial C-terminal region of ompB with a C-terminal 6xHis tag and >90% purity. However, outer membrane proteins require specific folding pathways and often need proper membrane integration for full functionality. The C-terminal His-tag may interfere with the native protein structure, particularly if this region is important for membrane anchoring or protein-protein interactions. Since activity is unverified, the protein cannot be assumed to be correctly folded or bioactive without experimental validation of its structural integrity and potential function.
1. Antibody Development and Validation Studies
This recombinant OmpB fragment is suitable for use as an immunogen to generate antibodies specific to this region of R. conorii OmpB. The His-tag facilitates purification and screening. However, it is critical to note that the antibodies generated will be against the linear epitopes of this non-native, likely misfolded fragment. Their ability to recognize the correctly folded, full-length OmpB protein in the outer membrane of R. conoriiis uncertain and must be empirically validated. These antibodies are primarily tools for detecting the denatured protein in techniques like Western blotting.
2. Protein-Protein Interaction Studies Using His-Tag Affinity
This application requires caution. While the His-tag enables technical feasibility for pull-down assays, if the ompB fragment is misfolded or improperly structured, it may not interact physiologically with true binding partners. Outer membrane proteins require precise conformation for specific interactions. This application should only be pursued after confirming proper folding through biophysical characterization.
3. ELISA-Based Binding Assays
This application is feasible for technical development but has limitations for functional studies. The His-tag enables ELISA format development, but if ompB is misfolded, binding studies may not reflect biological reality. The assay may work for antibody detection, but requires validation against properly folded ompB for accurate biological interaction studies.
4. Biochemical Characterization and Stability Studies
This purified recombinant ompB protein is well-suited for basic biochemical and biophysical characterization. Researchers can analyze its thermal stability, aggregation state, and solution behavior using techniques like dynamic light scattering and differential scanning calorimetry. This application is valid and independent of the protein's native bioactivity, as it focuses on the intrinsic physical properties of the purified polypeptide itself.
Final Recommendation & Action Plan
Given that ompB is a bacterial protein expressed in a prokaryotic system, but as a partial fragment with a tag, recommend first performing validation studies: 1) Biophysical characterization (circular dichroism for secondary structure, analytical size-exclusion chromatography for oligomeric state) to assess folding quality; 2) If possible, comparison with full-length ompB or native protein from R. conorii; 3) Functional validation of membrane binding or interaction properties if applicable. Antibody development can proceed as the safest application. Avoid functional interaction studies until proper folding is confirmed. For reliable ompB research, include appropriate controls such as known binding partners when possible.
