MyPseudomonas

Precision Genome Engineering Platform for Pseudomonas

Engineering Workflow

From target selection to a verified Pseudomonas mutant in five streamlined steps.

Our workflow combines cosmid-based homologous recombination, Red/ET recombineering, gusA blue-white screening, and the Iterative Marker Excision System (IMES) cassette for optional marker removal (Myronovskyi et al.). The cosmid containing the target region provides long genomic homology regions, while the IMES cassette is introduced using only short homology sequences. Following integration into Pseudomonas, blue-white screening helps identify double-crossover recombinants, which are subsequently verified by PCR. For marker-free engineering, the antibiotic resistance cassette can be excised using the φC31 integrase system (Myronovskyi et al.).

Pseudomonas mutants engineering - Cosmid manipulation Pseudomonas transformation Pseudomonas mutants blue-white screen Pseudomonas mutants engineering - marker excision Pseudomonas mutants plasmid curing

The Advantages of Our Platform

Products & Services

Genome-Wide Cosmids

Cosmid libraries covering full genomes with tiling overlap.

  • 25–40 kb inserts
  • End-sequenced clones
  • Replicate in E. coli

Deletion Kits

Complete gene deletion package for your Pseudomonas strain of interest.

  • Cosmid containing the chromosomal region of interest
  • IMES cassette (Apr / Tet)
  • Recombination strain
  • Helper vector for IMES cassette excision
  • Protocol and technical support

Custom Strain Engineering

Complete genome engineering service for your Pseudomonas strain.

  • Target analysis and strategy
  • Cosmid and IMES cassette construction
  • Single or multiple gene deletions
  • PCR and sequencing validation
  • Delivery of verified engineered strains