title: Allelic Exchange.
abstract: Methods used to understand the function of a gene/protein are one of the hallmarks of modern molecular genetics. The ability to genetically manipulate bacteria has become a fundamental tool in studying these organisms and while basic cloning has become a routine task in molecular biology laboratories, generating directed mutations can be a daunting task. This chapter describes the method of allelic exchange in Staphylococcus aureus using temperature-sensitive plasmids that have successfully produced a variety of chromosomal mutations, including in-frame deletions, insertion of antibiotic-resistance cassettes, and even single-nucleotide point mutations.
authors: Department of Pathology and Microbiology, Center for Staphylococcal Research, University of Nebraska Medical Center, Omaha, NE, 68198, USA.
authors: McKenzie K
authors: MK
authors: Lehman
authors: Department of Microbiology, Molecular Genetics and Immunology, The University of Kansas Medical Center, MSN 3029, 3901 Rainbow Boulevard, Kansas City, KS, 66160, USA.
authors: Jeffrey L
authors: JL
authors: Bose
authors: Department of Pathology and Microbiology, Center for Staphylococcal Research, University of Nebraska Medical Center, Omaha, NE, 68198, USA. kbayles@unmc.edu.
authors: Kenneth W
authors: KW
authors: Bayles
mesh_terms: D000483:Alleles; D003001:Cloning, Molecular / Q000379:methods*; D004352:Drug Resistance, Microbial / Q000235:genetics; D006801:Humans; D008967:Molecular Biology / Q000379:methods*; D016254:Mutagenesis, Insertional / Q000379:methods*; D010957:Plasmids; D011995:Recombination, Genetic; D013211:Staphylococcus aureus / Q000235:genetics* / Q000472:pathogenicity
keywords: Allelic exchange; Cloning; Homologous recombination; Mutation