This strain is a specialized "hyper-vesiculating" platform designed for the mass production of Outer Membrane Vesicles (OMVs). By disrupting both the structural anchoring of the cell envelope and the lipid transport machinery, the strain sheds high concentrations of vesicles into the culture medium.
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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Product Information
Product Overview
This strain is a specialized "hyper-vesiculating" platform designed for the mass production of Outer Membrane Vesicles (OMVs). By disrupting both the structural anchoring of the cell envelope and the lipid transport machinery, the strain sheds high concentrations of vesicles into the culture medium.
Target
Escherichia coli Nissle 1917
Applications
Ideal for producing OMV-based vaccines or drug delivery vehicles. It provides a significantly higher OMV yield compared to wild-type strains, reducing the cost and complexity of downstream purification.
Product Identity & Gene Details
Chassis Strain
Escherichia coli Nissle 1917
Target Gene(s)
tolR and mlaE
Gene Function
tolR: Encodes a protein in the Tol-Pal complex that links the inner and outer membranes; its absence causes "blebbing" of the outer membrane.
mlaE: Part of the Mla (Maintenance of Lipid Asymmetry) pathway; its deletion leads to an accumulation of phospholipids in the outer leaflet of the outer membrane, further destabilizing it.
Modification Type
Double gene knockout (Cell envelope integrity and lipid asymmetry)
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.
