Recombinant Rickettsia conorii Outer membrane protein A (ompA) is expressed in E. coli with an N-terminal 10xHis-SUMO tag and a C-terminal Myc tag. This partial protein covers amino acid region 1734-2021 and appears to achieve high purity levels exceeding 85% based on SDS-PAGE verification. The product is intended for research applications, providing what seems to be a dependable tool for scientists examining this particular ompA protein segment.
Outer membrane protein A (ompA) from Rickettsia conorii likely plays a critical role in bacterial cell membrane function. It contributes to structural integrity and may be important for interactions with host environments. Scientists frequently investigate this protein when studying pathogenicity and host-pathogen dynamics, which makes it an attractive research target in infectious disease and microbiology work.
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 ompA 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. However, the expressed fragment (1734-2021aa) represents only a partial sequence (288aa) of the full-length ompA, and contains dual tags (N-terminal 10xHis-SUMO and C-terminal Myc) that may interfere with native protein structure. Outer membrane proteins typically require specific folding pathways and membrane integration for proper function. While E. coli can express bacterial membrane proteins, the partial nature and tags may affect correct folding. Since activity is unverified, the protein cannot be assumed to be correctly folded or bioactive without experimental validation.
1. Antibody Development and Immunoassay Research
This recombinant OmpA fragment is suitable for use as an immunogen to generate polyclonal or monoclonal antibodies. The dual tags provide flexible options for purification and detection during antibody screening. The >85% purity is adequate for immunization protocols. However, it is crucial to understand that the antibodies generated will be against the linear epitopes of this non-native fragment. Their ability to recognize the full-length, correctly folded, and membrane-embedded OmpA protein in Rickettsia conoriiis very low and requires rigorous validation. The primary utility of these antibodies will be for detecting the denatured protein (e.g., in Western blots) or the recombinant immunogen itself in ELISA.
2. Protein-Protein Interaction Studies Using Tag-Assisted Pull-Down Assays
This application requires caution. While the dual tags enable technical feasibility for pull-down assays, if ompA is misfolded or improperly structured due to the partial sequence and tags, it may not interact physiologically with true binding partners. Outer membrane proteins require precise conformation for specific interactions with host cell receptors or other bacterial components. This application should only be pursued after confirming proper folding through biophysical characterization.
3. ELISA-Based Binding and Recognition Studies
This application is feasible for detection purposes but has limitations for functional studies. The tags enable technical development of ELISA formats, but if ompA is misfolded, binding studies may not reflect biological reality. The assay may work for antibody detection, but requires validation against properly folded ompA for accurate biological interaction studies.
4. Comparative Immunoblot Analysis of Rickettsial Proteins
This application is appropriate as a detection tool. The recombinant ompA can serve as a positive control and molecular weight standard for Western blot analysis. The dual tags provide multiple detection options. However, the migration pattern may differ from native ompA due to the tags and potential folding differences.
Final Recommendation & Action Plan
Given that ompA is a bacterial protein expressed in a prokaryotic system, but with a partial sequence and tags, recommend first performing validation studies: 1) Biophysical characterization (size-exclusion chromatography, circular dichroism) to assess folding state; 2) If possible, comparison with full-length ompA or native protein from Rickettsia cultures; 3) Functional validation of membrane binding or adhesion properties if applicable. Antibody development and use as a detection standard can proceed as relatively safe applications. For interaction studies, await proper folding confirmation and include appropriate controls such as known binding partners. Always validate findings with native ompA when possible.
