title: Construction of mouse 129/Ola BAC library for targeting experiments using E14 embryonic stem cells.
abstract: Gene targeting is a powerful method of specifically modifying genes of interest. It has been most consistently successful in the 129 mouse strain, because the embryonic stem (ES) cells of 129 mice are relatively easy to culture. In gene-targeting experiments, the use of ES cell-derived genomic clones as a source of homology arms is desirable, because the genetic variation among mouse strains results in a reduced frequency of homologous recombination. In this study, we generated an arrayed mouse 129/Ola BAC library derived from E14.1 ES cells, one of the frequently used ES cell lines. More than 135,000 BAC clones with a mean insert size of 110 kb were isolated. This library is estimated to represent a 5.5-fold mouse genome coverage. The BAC clones can be screened within 2 days by PCR. Considering that all 8 loci so far examined are contained in this BAC library, we believe it will be a useful resource for gene targeting studies using E14 ES cells as well as for genome analysis.
authors.affiliation: Division of Basic Molecular Science and Molecular Medicine, Tokai University School of Medicine, Isehara, Kanagawa, Japan. masato@is.icc.u-tokai.ac.jp
authors.forename: Masato
authors.initials: M
authors.lastname: Ohtsuka
authors.forename: Kenta
authors.initials: K
authors.lastname: Ishii
authors.forename: Yukie Y
authors.initials: YY
authors.lastname: Kikuti
authors.forename: Takayuki
authors.initials: T
authors.lastname: Warita
authors.forename: Daisuke
authors.initials: D
authors.lastname: Suzuki
authors.forename: Masahiro
authors.initials: M
authors.lastname: Sato
authors.forename: Minoru
authors.initials: M
authors.lastname: Kimura
authors.forename: Hidetoshi
authors.initials: H
authors.lastname: Inoko
mesh_terms: D000818:Animals; D022202:Chromosomes, Artificial, Bacterial / Q000378:metabolism*; D004622:Embryo, Mammalian / Q000166:cytology*; D018390:Gene Targeting / Q000379:methods; D015698:Genomic Library; D051379:Mice; D008815:Mice, Inbred Strains; D013234:Stem Cells / Q000378:metabolism*