title: A genome-wide, end-sequenced 129Sv BAC library resource for targeting vector construction.
abstract: The majority of gene-targeting experiments in mice are performed in 129Sv-derived embryonic stem (ES) cell lines, which are generally considered to be more reliable at colonizing the germ line than ES cells derived from other strains. Gene targeting is reliant on homologous recombination of a targeting vector with the host ES cell genome. The efficiency of recombination is affected by many factors, including the isogenicity (H. te Riele et al., 1992, Proc. Natl. Acad. Sci. USA 89, 5128-5132) and the length of homologous sequence of the targeting vector and the location of the target locus. Here we describe the double-end sequencing and mapping of 84,507 bacterial artificial chromosomes (BACs) generated from AB2.2 ES cell DNA (129S7/SvEvBrd-Hprtb-m2). We have aligned these BACs against the mouse genome and displayed them on the Ensembl genome browser, DAS: 129S7/AB2.2. This library has an average insert size of 110.68 kb and average depth of genome coverage of 3.63- and 1.24-fold across the autosomes and sex chromosomes, respectively. Over 97% of the mouse genome and 99.1% of Ensembl genes are covered by clones from this library. This publicly available BAC resource can be used for the rapid construction of targeting vectors via recombineering. Furthermore, we show that targeting vectors containing DNA recombineered from this BAC library can be used to target genes efficiently in several 129-derived ES cell lines.
authors.affiliation: The Wellcome Trust Sanger Institute, Hinxton, Cambridgeshire CB10 1SA, UK.
authors.forename: David J
authors.initials: DJ
authors.lastname: Adams
authors.forename: Michael A
authors.initials: MA
authors.lastname: Quail
authors.forename: Tony
authors.initials: T
authors.lastname: Cox
authors.forename: Louise
authors.initials: L
authors.lastname: van der Weyden
authors.forename: Barbara D
authors.initials: BD
authors.lastname: Gorick
authors.forename: Qin
authors.initials: Q
authors.lastname: Su
authors.forename: Wei-in
authors.initials: WI
authors.lastname: Chan
authors.forename: Rob
authors.initials: R
authors.lastname: Davies
authors.forename: James K
authors.initials: JK
authors.lastname: Bonfield
authors.forename: Frances
authors.initials: F
authors.lastname: Law
authors.forename: Sean
authors.initials: S
authors.lastname: Humphray
authors.forename: Bob
authors.initials: B
authors.lastname: Plumb
authors.forename: Pentao
authors.initials: P
authors.lastname: Liu
authors.forename: Jane
authors.initials: J
authors.lastname: Rogers
authors.forename: Allan
authors.initials: A
authors.lastname: Bradley
mesh_terms: D000818:Animals; D001483:Base Sequence; D002874:Chromosome Mapping; D022202:Chromosomes, Artificial, Bacterial / Q000235:genetics*; D015723:Gene Library; D018390:Gene Targeting / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics*; D023281:Genomics / Q000379:methods; D051379:Mice / Q000235:genetics*; D008969:Molecular Sequence Data; D020641:Polymorphism, Single Nucleotide / Q000235:genetics; D017422:Sequence Analysis, DNA; D013234:Stem Cells / Q000166:cytology