This strain is a "leaky" phenotype engineered to bypass the traditional need for mechanical cell lysis during protein harvesting. By removing a major structural anchor, the cell wall becomes porous, allowing internal products to diffuse into the surrounding broth.
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 "leaky" phenotype engineered to bypass the traditional need for mechanical cell lysis during protein harvesting. By removing a major structural anchor, the cell wall becomes porous, allowing internal products to diffuse into the surrounding broth.
Target
Escherichia coli Nissle 1917
Applications
Facilitates the extracellular production of large molecules like Fab fragments (antibodies). This simplifies the manufacturing process and protects sensitive proteins from the harsh conditions of industrial homogenization.
Product Identity & Gene Details
Chassis Strain
Escherichia coli Nissle 1917
Target Gene(s)
lpp
Gene Function
Encodes Braun's lipoprotein, which is the most abundant protein in E. coli and serves to tether the outer membrane to the underlying peptidoglycan layer.
Modification Type
Single gene knockout (Structural destabilization)
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.
