title: Bacterial artificial chromosome mutagenesis using recombineering.
abstract: Gene expression from bacterial artificial chromosome (BAC) clones has been demonstrated to facilitate physiologically relevant levels compared to viral and nonviral cDNA vectors. BACs are large enough to transfer intact genes in their native chromosomal setting together with flanking regulatory elements to provide all the signals for correct spatiotemporal gene expression. Until recently, the use of BACs for functional studies has been limited because their large size has inherently presented a major obstacle for introducing modifications using conventional genetic engineering strategies. The development of in vivo homologous recombination strategies based on recombineering in E. coli has helped resolve this problem by enabling facile engineering of high molecular weight BAC DNA without dependence on suitably placed restriction enzymes or cloning steps. These techniques have considerably expanded the possibilities for studying functional genetics using BACs in vitro and in vivo.
authors.affiliation: Department of Genetics and Genomic Sciences, Mount Sinai School of Medicine, New York, NY 10029, USA. kumaran.narayanan@sci.monash.edu.my
authors.forename: Kumaran
authors.initials: K
authors.lastname: Narayanan
authors.forename: Qingwen
authors.initials: Q
authors.lastname: Chen
mesh_terms: D000818:Animals; D022202:Chromosomes, Artificial, Bacterial; D004926:Escherichia coli / Q000235:genetics; D005818:Genetic Engineering / Q000379:methods*; D006801:Humans; D051379:Mice; D016296:Mutagenesis; D011995:Recombination, Genetic