A genome-wide, end-sequenced 129Sv BAC library resource for targeting vector construction

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 target...

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Veröffentlicht in:Genomics (San Diego, Calif.) Calif.), 2005-12, Vol.86 (6), p.753-758
Hauptverfasser: Adams, David J., Quail, Michael A., Cox, Tony, van der Weyden, Louise, Gorick, Barbara D., Su, Qin, Chan, Wei-in, Davies, Rob, Bonfield, James K., Law, Frances, Humphray, Sean, Plumb, Bob, Liu, Pentao, Rogers, Jane, Bradley, Allan
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container_issue 6
container_start_page 753
container_title Genomics (San Diego, Calif.)
container_volume 86
creator Adams, David J.
Quail, Michael A.
Cox, Tony
van der Weyden, Louise
Gorick, Barbara D.
Su, Qin
Chan, Wei-in
Davies, Rob
Bonfield, James K.
Law, Frances
Humphray, Sean
Plumb, Bob
Liu, Pentao
Rogers, Jane
Bradley, Allan
description 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- Hprt b-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.
doi_str_mv 10.1016/j.ygeno.2005.08.003
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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- Hprt b-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.</abstract><cop>San Diego, CA</cop><pub>Elsevier Inc</pub><pmid>16257172</pmid><doi>10.1016/j.ygeno.2005.08.003</doi><tpages>6</tpages><oa>free_for_read</oa></addata></record>
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subjects 129Sv
Animals
BAC library
Base Sequence
Biological and medical sciences
Chromosome Mapping
Chromosomes, Artificial, Bacterial - genetics
Fundamental and applied biological sciences. Psychology
Gene Library
Gene targeting
Gene Targeting - methods
Genes. Genome
Genetic Vectors - genetics
Genetics of eukaryotes. Biological and molecular evolution
Genomics - methods
Mice - genetics
Molecular and cellular biology
Molecular genetics
Molecular Sequence Data
Mouse genome
Polymorphism, Single Nucleotide - genetics
Sequence Analysis, DNA
Stem Cells - cytology
title A genome-wide, end-sequenced 129Sv BAC library resource for targeting vector construction
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