This strain is a classic "hyper-blebbing" model characterized by a significant loss of connection between the inner and outer bacterial membranes. It is primarily used in research to study membrane dynamics or to harvest large quantities of bacterial surface components.Escherichia coli Nissle 1917 tolR Mutant Are Highly Heterogeneous and Show Reduced Capacity for Epithelial Cell Interaction and Entry
For Research Use Only. Not intended for use in food manufacturing or medical procedures (diagnostics or therapeutics). Do Not Use in Humans.
| Product Information | |
|---|---|
| Product Overview | This strain is a classic "hyper-blebbing" model characterized by a significant loss of connection between the inner and outer bacterial membranes. It is primarily used in research to study membrane dynamics or to harvest large quantities of bacterial surface components.Escherichia coli Nissle 1917 tolR Mutant Are Highly Heterogeneous and Show Reduced Capacity for Epithelial Cell Interaction and Entry |
| Target | Escherichia coli Nissle 1917 |
| Applications | Excellent for rapid production of OMVs. The major advantage is the sheer volume of vesicles produced, though the strain may require more careful culturing due to its increased sensitivity to environmental stress. |
| Product Identity & Gene Details | |
|---|---|
| Chassis Strain | Escherichia coli Nissle 1917 |
| Target Gene(s) | tolR |
| Gene Function | A component of the Tol-Pal system, which uses the proton motive force to maintain the tension and positioning of the outer membrane relative to the cell wall. |
| Modification Type | Single gene knockout (Envelope structural integrity) |
| Genetic Architecture | |
|---|---|
| Promoter System | Native promoter |
| Selection Marker | Standard Antibiotic Marker or Markerless/Scarless (No resistance genes) |
| Delivery & Service Standards | |
|---|---|
| Deliverables | Glycerol Stocks |
| Shipping Conditions | Dry Ice Shipping |
| Lead Time | 4-8 weeks |
| Documents | Certificate of Analysis (COA) or Project Report |
| Verification | PCR Sequencing |
| Engineered Strain Recovery & Handling | |
|---|---|
| Immediate Post-Delivery Steps | 1. Transfer to Storage: Immediately transfer the cryovials to a -80°C ultra-low temperature freezer. 2. Avoid Fluctuations: Do not store the vials in a "frost-free" freezer, as the temperature cycles can damage the cell membranes of the engineered strains. |
| Strain Recovery Protocol | To ensure maximum viability, follow this standard recovery procedure: 1. Preparation: Prepare the appropriate selective agar medium (e.g., LB agar + specific antibiotic as listed in the COA). 2. Partial Thawing (The "Scratch" Method): * Place the cryovial on dry ice. Do not thaw the entire vial. Use a sterile inoculation loop or pipette tip to "scratch" a small amount of ice from the surface of the frozen stock. 3. Streaking: Streak the cells onto the selective agar plate to obtain single colonies. 4. Incubation: Invert the plate and incubate at the specified temperature (e.g., 37°C for E. coli or 30°C for S. cerevisiae) for 16-24 hours. 5. Subculturing: Pick a single, well-isolated colony to inoculate into liquid media for your experiments. |
| Usage Precautions & Best Practices | |
|---|---|
| Antibiotic Pressure | 1. Maintenance: For strains containing plasmids or specific resistance markers, always use the correct concentration of antibiotics in both solid and liquid media to prevent plasmid loss or contamination. 2. Markerless Strains: If the strain is a "scarless" integrated knockout, antibiotic pressure is not required for maintenance, but periodic genotypic verification is recommended. |
| Induction Conditions | 1. If your strain uses an inducible promoter (e.g., T7/IPTG or Arabinose), ensure the culture reaches the optimal OD600 (typically 0.6-0.8) before adding the inducer. 2. Verify the optimal induction temperature, as engineered strains often produce better results at lower temperatures (e.g., 16-25°C) to prevent inclusion body formation. |
| Disposal | All waste (vials, tips, plates) must be autoclaved at 121°C for at least 20 minutes or chemically disinfected before disposal. |
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For Research Use Only. Not intended for use in food manufacturing or medical procedures (diagnostics or therapeutics). Do Not Use in Humans.
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