Recombinant Escherichia coli RNA polymerase sigma factor for flagellar operon (fliA) is expressed in E. coli (strain K12) and contains the complete protein sequence from amino acids 1 to 239. This product comes without any tags, which appears to preserve the native protein structure and function. Purity exceeds 85% as determined by SDS-PAGE analysis, making it appropriate for various experimental applications that demand high-quality reagents.
The sigma factor fliA in Escherichia coli seems crucial for initiating transcription of the flagellar operon. It likely plays an important role in regulating flagellum assembly—a key component for bacterial movement. This protein may be essential for studying how bacteria move and how gene expression is controlled in prokaryotes, potentially offering insights into the intricate mechanisms behind bacterial adaptation and survival.
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. In Vitro Transcription Assays for Flagellar Gene Expression Studies
This recombinant FliA sigma factor works well in reconstituted in vitro transcription systems for studying flagellar operon gene regulation in E. coli. Scientists can pair this protein with core RNA polymerase to examine promoter specificity and how transcription starts at FliA-dependent promoters. The system should allow researchers to investigate how FliA guides RNA polymerase to specific flagellar genes while analyzing transcriptional regulation mechanisms. These assays might help clarify the molecular basis of flagellar gene expression control in bacterial motility research.
2. Protein-DNA Interaction Studies
Scientists can use the recombinant FliA protein in electrophoretic mobility shift assays (EMSA) or surface plasmon resonance experiments to characterize how it binds DNA with flagellar promoter sequences. These studies may help identify specific DNA recognition elements and binding affinities linked to FliA-dependent transcription. Researchers can map promoter regions and investigate sequence-specific interactions that appear to govern flagellar operon regulation. The tag-free design of the protein ensures minimal interference with native protein-DNA interactions.
3. Biochemical Characterization and Structural Studies
This purified FliA protein serves as a useful tool for detailed biochemical analysis, including protein folding studies, stability assays, and structural characterization techniques like circular dichroism spectroscopy or X-ray crystallography. The high purity level (>85%) makes it suitable for biophysical studies aimed at understanding the molecular structure and conformational properties of this sigma factor. Scientists can investigate protein stability under different conditions and examine structural features that may contribute to its specific regulatory functions.
4. Antibody Development and Validation
The recombinant FliA protein can serve as an antigen for generating specific antibodies against the E. coli flagellar sigma factor. These antibodies would likely be valuable research tools for immunoblotting, immunoprecipitation, and immunofluorescence studies investigating flagellar regulation in bacterial cells. The full-length protein expression (1-239aa) suggests that antibodies generated would recognize the complete range of potential epitopes present in the native protein. Such antibodies might facilitate further research into FliA localization and expression patterns in E. coli.
5. Comparative Sigma Factor Studies
This recombinant FliA can be used in comparative biochemical studies alongside other bacterial sigma factors to understand the specificity and functional differences between various transcriptional regulators. Scientists can examine how different sigma factors compete for core RNA polymerase binding or investigate the evolutionary relationships between flagellar regulatory systems across bacterial species. Having purified FliA available enables direct comparison of binding kinetics, promoter preferences, and regulatory mechanisms with other well-characterized sigma factors.
