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Creative Biogene leverages extensive molecular biology expertise and a mature microbial manipulation platform to provide Recombineering-based genome editing services across diverse microbes. Our capabilities include point mutations, insertions, gene knockouts, and metabolic pathway engineering for Gram-negative species such as E. coli, Enterobacter, Salmonella, and Pseudomonas. From design to validation, we deliver rapid, scarless genome modifications through a fully integrated, end-to-end workflow tailored to client needs.
Recombineering is based on recombinase-mediated homologous recombination, where exogenous DNA exchanges with host chromosomal sequences under the action of enzymes such as RecA, RecET, or the λ Red system. Artificial systems like λ Red (Exo, Beta, Gam) and RecET greatly enhance recombination efficiency:
These components work together to enable precise DNA integration, supporting single-base edits, fragment replacements, or reporter insertions. Compared with conventional targeting or transposon methods, Recombineering achieves higher efficiency, accurate locus targeting, and avoids random insertions.
In microbial genome editing, gene fragments with homologous arms are delivered via suicide vectors through electroporation or conjugation. Recombinants are selected under antibiotics, and subsequent counter-selection (e.g., sacB, rpsL, galK) removes the marker cassette, resulting in scarless, precise modifications. The final constructs are confirmed by PCR and sequencing, making this method ideal for metabolic engineering, functional genomics, and strain optimization.
Creative Biogene integrates both RecET and λ Red systems for high-efficiency genome editing across diverse hosts. Our experience has optimized key parameters—including electroporation conditions, homologous arm length, marker design, and screening strategies—to ensure reproducible success.
Figure 1. Overview of recombineering-based microbial genome editing workflow. (Fels U, et al. 2020)
Our technology supports:
All experiments follow standardized workflows, with high-throughput PCR screening and sequencing confirmation to ensure strain stability and genetic clarity.
1Design & Construction of Homologous Recombination Plasmids: Design upstream and downstream homology arms (40–1000 bp) and clone target sequences into suicide vectors or FRT/loxP plasmids with selectable markers.
2Preparation of Rec System Host Strains: Introduce Rec or λ Red plasmids into host bacteria, verify colonies via PCR, and prepare competent cells for recombination.
3Homologous Recombination: Transform recombination fragments into Rec-positive competent cells, induce recombinase expression, and select positive clones by PCR and sequencing.
4Marker Removal & Verification: Introduce FLP recombinase plasmids to excise selection markers, achieving scarless mutants. Confirm modifications using PCR, Southern blot, or sequencing.
5Final Strain Validation: Assess physiological phenotype, metabolic profile, and growth curves to ensure no off-target effects. Deliver validated, engineered strains with a full report.
With years of experience in microbial genetics and molecular engineering, Creative Biogene is a leading provider of microbial genome editing services. Our comprehensive recombinase platform, expert scientific team, and rigorous quality management system deliver custom solutions from target design to final edited strain delivery.
Whether your goal is gene function analysis, metabolic product optimization, or construction of novel synthetic microbial strains, Creative Biogene provides efficient, precise, and verifiable technical support. Contact us today to explore how our Recombineering services can accelerate your microbial genome engineering projects.
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