Molecular Life Sciences

Living Edition
| Editors: Robert D. Wells, Judith S. Bond, Judith Klinman, Bettie Sue Siler Masters, Ellis Bell


Living reference work entry



Recombineering is a method for constructing recombinant DNA molecules in vivo, using homologous recombination in E. coli or other cells (Sawitzke et al. 2007; Sharan et al. 2009). In this approach, linear dsDNA molecules introduced into the cell undergo recombination at homologous sequences to generate novel sequence combinations or to insert DNA into the chromosome. The cells used in the method express three bacteriophage proteins that (1) degrade the 5′-end of one strand of the dsDNA, (2) bind the resulting single-stranded DNA and promote its annealing with complementary DNA, and (3) protect the linear DNA from degradation by the E. coli RecBCD enzyme. The combined action of these three proteins brings about recombination of the linear DNA with homologous sequences in the cell’s chromosome or in other DNA in the cell, such as a plasmid or BAC (Fig. 1).


Bacteriophage Lambda dsDNA Molecule Introduce Point Mutation Linear dsDNA Lambda Exonuclease 
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  1. De Lay NR, Cronan JE (2006) Gene-specific random mutagenesis of Escherichia coli in vivo: isolation of temperature-sensitive mutations in the acyl carrier protein of fatty acid synthesis. J Bacteriol 188:287–296PubMedCentralPubMedCrossRefGoogle Scholar
  2. Muyrers JP, Zhang Y, Benes V, Testa G, Ansorge W, Stewart AF (2000) Point mutation of bacterial artificial chromosomes by ET recombination. EMBO Rep 1:239–243PubMedCentralPubMedCrossRefGoogle Scholar
  3. Sawitzke JA, Thomason LC, Costantino N, Bubunenko M, Datta S, Court DL (2007) Recombineering: in vivo genetic engineering in E. coli, S. enterica, and beyond. Methods Enzymol 421:171–199PubMedCrossRefGoogle Scholar
  4. Sharan SK, Thomason LC, Kuznetsov SG, Court DL (2009) Recombineering: a homologous recombination-based method of genetic engineering. Nat Protoc 4:206–223PubMedCentralPubMedCrossRefGoogle Scholar
  5. Yu D, Ellis HM, Lee EC, Jenkins NA, Copeland NG, Court DL (2000) An efficient recombination system for chromosome engineering in Escherichia coli. Proc Natl Acad Sci U S A 97:5978–5983PubMedCentralPubMedCrossRefGoogle Scholar

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© Springer Science+Business Media New York 2014

Authors and Affiliations

  1. 1.Department of Chemistry & BiochemistryUniversity of MarylandCollege ParkUSA