title: Recombining overlapping BACs into single large BACs.
abstract: BAC clones containing the entire genomic region of a gene including the long-range regulatory elements are very useful for gene functional analysis. However, large genes often span more than the insert of a BAC clone, and single BACs covering the entire region of interest are not available. Here, we describe a general system for linking two or more overlapping BACs into a single clone. Two rounds of homologous recombination are used. In the first, the BAC inserts are subcloned into the pBACLink vectors. In the second, the two BACs are combined together. Multiple BACs in a contig can be combined by alternating use of the pBACLInk vectors, resulting in several BAC clones containing as much of the genomic region of a gene as required. Such BACs can then be used in gene expression studies and/or gene therapy applications.
authors.affiliation: Department of Histology and Embryology, School of Medicine, University of Athens, 75 Mikras Asias Str., Athens, 11527, Greece, geokotz@med.uoa.gr.
authors.forename: George
authors.initials: G
authors.lastname: Kotzamanis
authors.forename: Athanassios
authors.initials: A
authors.lastname: Kotsinas
mesh_terms: D022202:Chromosomes, Artificial, Bacterial / Q000737:chemistry / Q000378:metabolism*; D003001:Cloning, Molecular / Q000379:methods*; D018274:Electroporation; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors / Q000737:chemistry / Q000378:metabolism*; D015698:Genomic Library; D059765:Homologous Recombination; D010957:Plasmids / Q000737:chemistry / Q000378:metabolism*; D019076:Transgenes