Establishment of an Efficient BAC Transgenesis Protocol and its Application to Functional Characterization of the Mouse Brachyury Locus

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Transgenesis using large DNA such as YAC or BAC has extended the range of applications in functional genomics. Here we describe an efficient BAC transgenesis protocol using a simple BAC DNA preparation method adopted from YAC DNA purification methods. This method allowed us to isolate BAC DNA from small scale culture of BAC-containing cells in sufficient quantity and purity for microinjection. More than 40 founders have been produced with linearized BAC DNA prepared by this method, and 85% of them contained intact BAC transgenes. In contrast, when circular BAC DNA was injected, an approximately three-fold reduction of transgene integration rate was observed and fewer intact transgene integrations were obtained. A line of transgenic mice carrying a 170-kb BAC clone generated in this way successfully rescued tail and embryonic lethality phenotypes of the mouse <i>Brachyury</i> (<i>T</i>) mutants, further demonstrating the utility of this method in functional analysis of the mouse genome.<br>

収録刊行物

  • Experimental animals

    Experimental animals 53(4), 311-320, 2004-07

    公益社団法人 日本実験動物学会

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各種コード

  • NII論文ID(NAID)
    10013441566
  • NII書誌ID(NCID)
    AA11032321
  • 本文言語コード
    ENG
  • 資料種別
    ART
  • ISSN
    13411357
  • NDL 記事登録ID
    7016984
  • NDL 雑誌分類
    ZS7(科学技術--医学)
  • NDL 請求記号
    Z54-H752
  • データ提供元
    CJP書誌  NDL  J-STAGE 
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