Phage Lamada Red Recombination System Construction

Genetic recombination is defined as the rearrangement of genes or parts of genes. The genetic recombination process enhances host-encoded recombinant proteins with phage-encoded recombinant proteins that promote specific recombination pathways. Throughout evolution, the acquisition of "new" genes and the rearrangement of "old" genes are driven by genetic recombination between phage and bacterial genomes. In Escherichia coli, genetic recombination proceeds "cautiously" through carefully regulated mechanisms. While genetic recombination may be very active in repairing DNA damage or promoting genetic diversity during times of stress, uncontrolled recombination can lead to genomic instability. Phage lambda infects E. coli and induces a significantly increased and altered rate of genetic recombination as part of the infection cycle. This relatively "static" recombination mechanism changed dramatically when E. coli was infected with the temperate phage lambda. Each lambda offspring undergoes at least one recombination event, even though they possess a tiny chromosome only 50 kb in length.

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Recombinant phages can also be engineered to contain fluorescent and bioluminescent reporter genes, enabling their use in tracking, detection, and biosensing applications. Our researchers have developed a recombination phage platform technology that can be easily and precisely engineered for specific phage applications.

• Genetic recombination in phage lamada
• Expression of Lambda Red recombination genes
• Lambda red recombineering design
• Lambda red recombineering construction in E.coli

Key Components of the Lambda Red Recombineering System

The lambda red recombination system has three components: Exo, Beta, and Gam. All three are required for recombination with dsDNA substrates; however, when using ssDNA substrates to generate modifications, only Beta is required.

• Gam: Gam prevents endogenous RecBCD and SbcCD nucleases from digesting linear DNA introduced into E. coli.
• Exo: Exo is a 5'→3' dsDNA-dependent exonuclease. Exo will degrade linear dsDNA starting from the 5' end and produce two possible products: 1) a partial dsDNA duplex with single-stranded 3' overhangs or 2) ssDNA in which the entire complementary strand is degraded if the dsDNA is short enough.
• Beta: Beta protects the ssDNA created by Exo and promotes its annealing to the complementary ssDNA target in the cell. Only Beta expression is required for recombination with ssDNA oligonucleotide substrates.

Workflow of Lambda Red Recombineering Construction

• Substrate DNA Design and Generation
• Expression of the λ red recombinant gene
• Electroporation of substrate DNA and growth of bacteria
• Selection and confirmation of recombinant clones
• Lambda red recombineering construction

Lambda Red Recombineering Construction Application

• Eliminate bacteria in biomedical, industrial or ecological environments
• High sensitivity biosensors for detection of specific bacteria or environmental conditions
• Easy to lyse recombinant protein-producing bacteria for rapid extraction

Creative Biolabs can meet the needs of customers by providing construction services of phage λ red recombination system 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.

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