ArticleMolecular biotechnology2006
An improved recombineering approach by adding RecA to lambda Red recombination.
Article in Molecular biotechnology, 2006. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 82 papers.
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Who cites it
82 citing papers in PubMed, 140 citations in OpenAlex.
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- Efficient markerless integration of genes in the chromosome of probiotic E. coli Nissle 1917 by bacterial conjugation.Microbial biotechnology · 2022Article
- HR-Bac, a toolbox based on homologous recombination for expression, screening and production of multiprotein complexes using the baculovirus expression system.Scientific reports · 2022Article
- A versatile genetic engineering toolkit for E. coli based on CRISPR-prime editing.Nature communications · 2021Article
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- Mutational Analysis of Redβ Single Strand Annealing Protein: Roles of the 14 Lysine Residues in DNA Binding and Recombination In Vivo.International journal of molecular sciences · 2021Article
- Oligomeric complexes formed by Redβ single strand annealing protein in its different DNA bound states.Nucleic acids research · 2021Article
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- The mole genome reveals regulatory rearrangements associated with adaptive intersexuality.Science (New York, N.Y.) · 2020Article
22 more citing papers are in PubMed but not listed here.
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Authors and funding
9 authors at 3 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Recombineering is the use of homologous recombination in Escherichia coli for DNA engineering. Of several approaches, use of the lambda phage Red operon is emerging as the most reliable and flexible. The Red operon includes three components: Redalpha, a 5' to 3' exonuclease, Redbeta, an annealing protein, and Redgamma, an inhibitor of the major E. coli exonuclease and recombination complex, RecBCD. Most E. coli cloning hosts are recA deficient to eliminate recombination and therefore enhance the stability of cloned DNAs. However, loss of RecA also impairs general cellular integrity. Here we report that transient RecA co-expression enhances the total number of successful recombinations in bacterial artificial chromosomes (BACs), mostly because the E. coli host is more able to survive the stresses of DNA transformation procedures. We combined this practical improvement with the advantages of a temperature-sensitive version of the low copy pSC101 plasmid to develop a protocol that is convenient and more efficient than any recombineering procedure, for use of either double- or single-stranded DNA, published to date.
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