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FISH analysis of 142 EGFP transgene integration sites into the mouse genome
Authors:Nakanishi Tomoko  Kuroiwa Asato  Yamada Shuichi  Isotani Ayako  Yamashita Atsuko  Tairaka Ai  Hayashi Tamon  Takagi Tatsuya  Ikawa Masahito  Matsuda Yoichi  Okabe Masaru
Affiliation:Genome Information Research Center, Osaka University, Yamadaoka 3-1, Suita Osaka, 565-0871, Japan.
Abstract:Production of transgenic animals is an important technique for studying various biological processes. However, whether the integration of a particular transgene occurs randomly in the mouse genome has not been determined. Analysis by fluorescence in situ hybridization of the integration sites of the 142 EGFP (a mutant of green fluorescent protein) transgenic lines that we produced showed that the transgenes had become incorporated into every mouse chromosome. A single integration site was observed in 82.4% of the lines. The concomitant integrations of transgene into two different loci were observed in 15 cases (10.6%). In 3 cases, the transgenic founder mice showed chimerism in integration sites (2.1%). Chromosomal translocation was observed in 7 cases (4.9%). Moreover, when we statistically analyzed the transgene integration sites of these mouse lines, they were shown to distribute unevenly throughout the genome. This is the first report to analyze the transgene integration sites by producing more than 100 transgenic mouse lines.
Keywords:FISH analysis   transgene integration   green fluorescent protein   transgenic mouse
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