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Phage Recombineering of Electroporated DNA

Bacteriophage recombineering of electroporated DNA (BRED) for simple and efficient construction of targeted phage mutants. BRED can be used to construct untagged deletions of essential and non-essential genes, in-frame deletions, point and nonsense mutations, addition of gene tags, and precise insertion of foreign genes. This technique is applicable to mycobacteriophages and should be generally applicable to phages of other hosts for which recombinant systems are available. It involves the co-electroporation of recombination substrates (i.e., phage DNA and double-stranded DNA (dsDNA)) into electrocompetent bacterial cells carrying plasmids encoding proteins that promote high levels of homologous recombination, such as RecE/RecT-like proteins. By using this method, modified phages can be obtained at high frequency (10% to 15%), allowing screening of putative mutants by small amounts of PCR without further selection. This technique requires a highly competent bacterial host.

Fig 1:Bacteriophage recombineering of electroporated DNAFig 1. Bacteriophage recombineering of electroporated DNA (Pires D P, et al. 2016)

Our Services

Creative Biolabs successfully obtained phage genetic modification using bacteriophage recombineering of electroporated DNA. With the accumulation of technology and resources, the reconstruction of the microbial genome is to realize the research and utilization of genes by interrupting, inserting, or deleting genes in the microbial genome.

  • Phage recombineering design
  • Phage screening and isolation
  • Construction of phage mutants defective in essential genes
  • Use of BRED to construct internal deletions of phage gene
  • Mutants generated have in-frame deletions in phage gene
  • Recombineering phage production

Fast Project Execution

We are months ahead of our competitors. We start projects within a day of client approval and complete pilot studies within 15-20 days. Subsequent large-scale phage recombineering takes 5-10 days, and product isolation is usually completed after 2-3 days. Our typical project timeline from start to finish is five weeks.

  • Project start time: 0 - 1 day
  • Pilot study: 15 - 20 days
  • Large scale recombineering: 5 - 10 days
  • Product quarantine: 2 - 3 days
  • Final Analysis: 2 - 3 days
  • Delivery: 1 - 2 working days

Fig 2: Workflow of bacteriophage recombineering

Advantages of Recombinant Technology

  • Ease of subcloning and modification of microorganisms
  • Scientists with experience in gene editing and immuno-oncology
  • Fast, simple, efficient and flexible
  • Independent of restriction sites
  • No size limits
  • Exact variation

Creative Biolabs can meet the needs of customers by providing bacteriophage recombineering of electroporated DNA services on time and on budget. We have in-depth knowledge and experience of the tools and processes involved in the phage projects. Our skilled and dedicated scientific researchers ensure that the most suitable methods and techniques are selected for your project. If necessary, please feel free to contact us.

Reference

  • Pires D P, Cleto S, Sillankorva S, et al. Genetically engineered phages: a review of advances over the last decade[J]. Microbiology and Molecular Biology Reviews, 2016, 80(3): 523-543.

Please kindly note that our services can only be used to support research purposes (Not for clinical use).

Biophage Technology

Creative Biolabs is a globally recognized phage company. Creative Biolabs is committed to providing researchers with the most reliable service and the most competitive price.

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