This is a therapeutic delivery strain engineered to secrete ClyA-fusion proteins using the bacteria's natural pore-forming protein as a "shuttle." It is specifically designed to export therapeutic payloads directly into the surrounding environment, such as a tumor microenvironment.
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 is a therapeutic delivery strain engineered to secrete ClyA-fusion proteins using the bacteria's natural pore-forming protein as a "shuttle." It is specifically designed to export therapeutic payloads directly into the surrounding environment, such as a tumor microenvironment.
Target
Escherichia coli Nissle 1917
Applications
Used in bacterial-mediated cancer therapy to deliver anti-tumor toxins or antigens. The advantage is that it utilizes an existing secretion pathway to "pump" drugs out of the bacteria and into target cells.
Product Identity & Gene Details
Chassis Strain
Escherichia coli Nissle 1917
Target Gene(s)
ClyA (HlyE) and Hy
Gene Function
ClyA (HlyE): A pore-forming cytotoxin that E. coli naturally exports via outer membrane vesicles.
Hy: Typically stands for a "Hybrid" or "Hyper" variant, or a specific fusion partner (like a specific antigen) attached to the ClyA carrier.
Modification Type
Recombinant protein expression (Secretion system)
Genetic Architecture
Promoter System
Constitutive or Inducible
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.
