A genome-wide, end-sequenced 129Sv BAC library resource for targeting vector construction
A genome-wide, end-sequenced 129Sv BAC library resource for targeting vector construction
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DOI：
10.1016/j.ygeno.2005.08.003
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发表时间：
2005-12-01
期刊：
影响因子：
4.4
通讯作者：
Bradley, A
中科院分区：
文献类型：
--
作者：
Adams, DJ;Quail, MA;Bradley, A
小鼠中的大多数基因靶向实验都是在129 Sv来源的胚胎干（ES）细胞系中进行的，通常认为这些细胞系比其他菌株来源的ES细胞在生殖系定殖方面更可靠。基因靶向依赖于靶向载体与宿主ES细胞基因组的同源重组。重组效率受许多因素影响，包括同源性（H。te Riele等人，1992，Proc.Natl. Acad. Sci. USA 89，5128-5132）和靶向载体的同源序列的长度以及靶基因座的位置。在这里，我们描述了从AB2.2 ES细胞DNA（129 S7/SvEvBrd-Hprt（b-m2））产生的84，507个细菌人工染色体（BAC）的双末端测序和作图。我们已经将这些BAC与小鼠基因组进行了比对，并将它们展示在Ensembl基因组浏览器DAS：129 S7/AB2.2上。该文库的平均插入片段大小为110.68 kb，在常染色体和性染色体上的平均基因组覆盖深度分别为3.63倍和1.24倍。超过97%的小鼠基因组和99.1%的Ensembl基因被来自该文库的克隆覆盖。这种公众可获得的BAC资源可用于通过重组工程快速构建靶向载体。此外，我们表明，靶向载体含有DNA重组从这个BAC库可以用来有效地在几个129衍生的ES细胞系的靶向基因。(c)2005年爱思唯尔公司All rights reserved.
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 oil 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. (c) 2005 Elsevier Inc. All rights reserved.