FISH analysis of 142 EGFP transgene integration sites into the mouse genome

被引:124
作者
Nakanishi, T
Kuroiwa, A
Yamada, S
Isotani, A
Yamashita, A
Tairaka, A
Hayashi, T
Takagi, T
Ikawa, M
Matsuda, Y
Okabe, M
机构
[1] Osaka Univ, Genome Informat Res Ctr, Suita, Osaka 5650871, Japan
[2] Hokkaido Univ, Ctr Adv Sci & Technol, Kita Ku, Sapporo, Hokkaido 0600810, Japan
[3] Program Promot Basic Res Act Innovat Biosci, Tokyo, Japan
[4] Osaka Univ, Grad Sch Pharmaceut Sci, Suita, Osaka 5650871, Japan
基金
日本学术振兴会;
关键词
FISH analysis; transgene integration; green fluorescent protein; transgenic mouse;
D O I
10.1006/geno.2002.7008
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
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.
引用
收藏
页码:564 / 574
页数:11
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