λ‐Red Genetic Engineering in Salmonella enterica serovar Typhimurium

Abstract The use of the recombination system from bacteriophage lambda, λ‐Red, allows for PCR‐generated fragments to be targeted to specific chromosomal locations in sequenced genomes. A minimal region of homology of 30 to 50 bases flanking the fragment to be inserted is all that is required for targeted mutagenesis. Procedures for creating specific insertions, deletions, and site‐directed changes are described.

[1]  A. Stewart,et al.  Techniques: Recombinogenic engineering--new options for cloning and manipulating DNA. , 2001, Trends in biochemical sciences.

[2]  K. Murphy,et al.  Lambda Red-mediated recombinogenic engineering of enterohemorrhagic and enteropathogenic E. coli , 2003, BMC Molecular Biology.

[3]  D. Court,et al.  A highly efficient Escherichia coli-based chromosome engineering system adapted for recombinogenic targeting and subcloning of BAC DNA. , 2001, Genomics.

[4]  A. Yamaguchi,et al.  Transport of divalent cations with tetracycline as mediated by the transposon Tn10-encoded tetracycline resistance protein. , 1990, The Journal of biological chemistry.

[5]  B. Wanner,et al.  One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[6]  A. Poteete What makes the bacteriophage λ Red system useful for genetic engineering: molecular mechanism and biological function , 2001 .

[7]  K. Murphy,et al.  PCR-mediated gene replacement in Escherichia coli. , 2000, Gene.

[8]  D. W. Daniels,et al.  Promoter mutations affecting divergent transcription in the Tn10 tetracycline resistance determinant. , 1985, Journal of molecular biology.

[9]  A. Kuzminov,et al.  Annealing vs. Invasion in Phage λ Recombination , 1997 .

[10]  S. Rubino,et al.  Epitope tagging of chromosomal genes in Salmonella , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[11]  D. Court,et al.  Genetic engineering using homologous recombination. , 2002, Annual review of genetics.

[12]  C. Wolk,et al.  Use of a conditionally lethal gene in Anabaena sp. strain PCC 7120 to select for double recombinants and to entrap insertion sequences , 1990, Journal of bacteriology.

[13]  R. Kolodner,et al.  Homologous pairing proteins encoded by the Escherichia coii recE and recT genes , 1994, Molecular microbiology.

[14]  J. Ghigo,et al.  A rapid and simple method for inactivating chromosomal genes in Yersinia. , 2003, FEMS immunology and medical microbiology.

[15]  D. Court,et al.  An efficient recombination system for chromosome engineering in Escherichia coli. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[16]  J. W. Little An exonuclease induced by bacteriophage lambda. II. Nature of the enzymatic reaction. , 1967, The Journal of biological chemistry.

[17]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[18]  J. Way,et al.  New Tn10 derivatives for transposon mutagenesis and for construction of lacZ operon fusions by transposition. , 1984, Gene.

[19]  Y. Sakaki,et al.  The gamma protein specified by bacteriophage gamma. Structure and inhibitory activity for the recBC enzyme of Escherichia coli. , 1975, The Journal of biological chemistry.

[20]  G. Reddy,et al.  The beta protein of phage lambda promotes strand exchange. , 1998, Journal of molecular biology.

[21]  Stanley Maloy,et al.  Selection for loss of tetracycline resistance by Escherichia coli , 1981, Journal of bacteriology.

[22]  Nancy A. Jenkins,et al.  Recombineering: a powerful new tool for mouse functional genomics , 2001, Nature Reviews Genetics.

[23]  A. Clark,et al.  Biochemical and genetic studies of recombination proficiency in Escherichia coli. II. Rec+ revertants caused by indirect suppression of rec- mutations. , 1970, Proceedings of the National Academy of Sciences of the United States of America.

[24]  K. Murphy,et al.  Lambda Gam protein inhibits the helicase and chi-stimulated recombination activities of Escherichia coli RecBCD enzyme , 1991, Journal of bacteriology.

[25]  N. Copeland,et al.  A highly efficient recombineering-based method for generating conditional knockout mutations. , 2003, Genome research.

[26]  Frank Buchholz,et al.  A new logic for DNA engineering using recombination in Escherichia coli , 1998, Nature Genetics.

[27]  A. Nadkarni,et al.  Chromosomal duplications and cointegrates generated by the bacteriophage lamdba Red system in Escherichia coli K-12 , 2004, BMC Molecular Biology.

[28]  K. Hughes,et al.  Genetic Transplantation: Salmonella enterica Serovar Typhimurium as a Host To Study Sigma Factor and Anti-Sigma Factor Interactions in GeneticallyIntractable Systems , 2006, Journal of bacteriology.