Recombinant Clostridium botulinum UPF0059 membrane protein CLI_0469 (CLI_0469)
Description
Production and Purification
The recombinant protein is synthesized in E. coli using standard prokaryotic expression systems. Critical steps include:
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Cloning: Insertion of the CLI_0469 gene into a plasmid vector with a His tag.
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Induction: IPTG-mediated expression optimization for yield.
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Purification: Nickel-affinity chromatography (via His tag) followed by buffer exchange.
Commercial Specifications (based on vendor data):
| Parameter | Value | Source |
|---|---|---|
| Storage Buffer | Tris-based buffer, 50% glycerol, pH optimized for stability | |
| Storage Conditions | -20°C (long-term), 4°C (short-term) | |
| Purity | >90% (SDS-PAGE verified) |
Research Applications
While limited peer-reviewed studies exist, CLI_0469 is primarily used in:
Antibody Production and ELISA Assays
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Antigen Source: Serves as a recombinant antigen for generating polyclonal or monoclonal antibodies.
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ELISA Kits: Commercially available as a coated antigen in immunoassays (e.g., CBM15’s ELISA kit).
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Example Protocol:
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Coat plates with CLI_0469 (50 µg/mL).
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Block with casein buffer.
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Incubate with primary antibodies (e.g., anti-C. botulinum sera).
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Detect using HRP-conjugated secondary antibodies.
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| Application | Purpose | Source |
|---|---|---|
| Serological Studies | Detecting anti-C. botulinum antibodies in sera | |
| Protein Interaction | Identifying binding partners via pull-down assays |
Limitations and Knowledge Gaps
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Functional Annotation: No published studies confirm its biological role in C. botulinum.
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Toxicity Data: No safety assessments or bioactivity reports available.
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Sequence Homology: Limited alignment with characterized proteins beyond UPF0059 family.
Table 1: Amino Acid Sequence Highlights
| Region | Residues | Function (Predicted) |
|---|---|---|
| N-Terminal | 1–50 (MEVQELFLLALAISLDAFGVILCIGINKGITLKSSIIFVFSFGFFQFFLSFLGGYIGTIF) | Membrane insertion signal? |
| C-Terminal | 134–183 (...SYINNLFYLFMSSLFMGLIATIICSLGIILSKYIKKISIISSYADYIGGIILILFGLKM LFF) | Cytosolic domain or binding site |
Product Specs
Note: While we prioritize shipping the format currently in stock, please specify your format preference in order remarks for customized preparation.
Note: Standard shipping includes blue ice packs. Dry ice shipping requires advance notification and incurs additional charges.
The specific tag type is determined during production. If you require a specific tag, please inform us; we will prioritize its development.
Target Background
This protein likely functions as a manganese efflux pump.
KEGG: cbf:CLI_0469
