Recombineering: Genetic Engineering in Bacteria Using Homologous Recombination

@article{Thomason2007RecombineeringGE,
  title={Recombineering: Genetic Engineering in Bacteria Using Homologous Recombination},
  author={Lynn C. Thomason and Donald L. Court and Mikail Bubunenko and Nina Costantino and Helen R. Wilson and Simanti Datta and Amos B. Oppenheim},
  journal={Current Protocols in Molecular Biology},
  year={2007},
  volume={78}
}
The bacterial chromosome and plasmids can be engineered in vivo by homologous recombination using PCR products and synthetic oligonucleotides as substrates. This is possible because bacteriophage‐encoded recombination functions efficiently to recombine sequences with homologies as short as 35 to 40 bases. This recombineering allows DNA sequences to be inserted or deleted without regard to location of restriction sites. This unit first describes preparation of electrocompetent cells expressing… 
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240 Citations
Highly Influential Citations
11
Background Citations
37
Methods Citations
72
Results Citations
1

240 Citations

Citation Type

Citation Type

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TLDR
Various recombineering strategies to promote different types of gene modifications, how they are best applied, and their possible pitfalls are discussed.
Recombineering: in vivo genetic engineering in E. coli, S. enterica, and beyond.
Recombineering: highly efficient in vivo genetic engineering using single-strand oligos.
Multicopy Plasmid Modification with Phage λ Red Recombineering
Phage recombinases and their applications.
  • K. Murphy
  • Biology
    Advances in virus research
  • 2012
A set of recombineering plasmids for gram-negative bacteria.
Genome modifications and cloning using a conjugally transferable recombineering system
Bacteriophage recombination systems and biotechnical applications
TLDR
Key steps of the molecular mechanisms involving phage recombination functions and their application to molecular engineering, the novel exploitations of the PY54-derived recombination system, and its application to the development of new DNA vectors are discussed.
Use of Recombination-Mediated Genetic Engineering for Construction of Rescue Human Cytomegalovirus Bacterial Artificial Chromosome Clones
TLDR
Construction of rescue clones using gap repair cloning is highly efficient and provides a novel use of the homologous recombination-based method in E. coli for molecular cloning, known colloquially as recombineering, when rescuing large BAC deletions.
Enhancement of RecET-mediated in vivo linear DNA assembly by a xonA mutation
TLDR
Deletion of the E. coli xonA gene, encoding Exonuclease I, a 3’→5’ single-strand DNA ex onuclease, substantially improves the efficiency of in vivo linear DNA assembly for both systems and provides a significant improvement to previously reported in vivolinear DNA assembly technologies.
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