Establishment of an Efficient BAC Transgenesis Protocol and its Application to Functional Characterization of the Mouse Brachyury Locus
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概要
- 論文の詳細を見る
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 Brachyury (T) mutants, further demonstrating the utility of this method in functional analysis of the mouse genome.
- 社団法人 日本実験動物学会の論文
著者
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Suzuki Misao
Center For Animal Resources And Development Kumamoto Univ.
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Suzuki Misao
Center For Animal Research And Development Kumamoto University
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Katoh Hideki
Experimental Animal Center Hamamatsu University School Of Medicine
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Yamamura Ken-ichi
Institute Of Molecular Embryology And Genetics Kumamoto University School Of Medicine
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Yamamura Ken-ichi
Institute For Medical Genetics Kumamoto University Medical School
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Yamamura Ken-ichi
Institute Of Molecular Embryology And Genetics Kumamoto University
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Abe Kuniya
Research & Development Team For Mammalian Cellular Dynamics Bioresource Center Riken Tsukuba Ins
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HAZAMA Masaaki
Department of Biological Sciences, Kyoto University Faculty of Medicine
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Hazama Masaaki
Department Of Biological Sciences Kyoto University Faculty Of Medicine
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Katoh Hideki
Experimental Animals Institute Hamamatsu University School Of Medicine
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