Datasheet

Escherichia coli Nissle 1917ΔmchBCDEFΔmcmIA (CAT#: LBGF-0126-GF27)

This strain is engineered to be deficient in the production and immunity of Microcin H47, a potent antibacterial peptide. It is used to study the role of bacteriocins in gut colonization or to create a "blank slate" for the insertion of new antimicrobial tools.

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 engineered to be deficient in the production and immunity of Microcin H47, a potent antibacterial peptide. It is used to study the role of bacteriocins in gut colonization or to create a "blank slate" for the insertion of new antimicrobial tools.
Target Escherichia coli Nissle 1917
Applications Useful for researchers wanting to "disarm" the natural competitive advantage of EcN. This allows for the precise study of how other engineered traits contribute to gut fitness without the interference of native antimicrobial production.
Product Identity & Gene Details
Chassis Strain Escherichia coli Nissle 1917
Target Gene(s) mchBCDEF and mcmIA
Gene Function mchBCDEF: A gene cluster responsible for the synthesis, processing, and transport of Microcin H47.

mcmIA: Genes providing immunity to the producer cell to prevent self-destruction by its own microcin.
Modification Type Multi-gene cluster deletion (Bacteriosin biosynthesis and immunity)
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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