PCR-based approaches for identification of multi-copy transgene integration sites in mouse genome  

作　　者：ZHAO Xudong DANG Suying LIANG Bin LEI Xia CHEN Zheng WANG Long YAN Lanzhen SUN Hantang FU Jiliang FEI Jian WANG Zhugang 

机构地区：[1]Shanghai Research Center for Model Organisms, Shanghai 201203, China [2]Laboratory of Genetic Engineering, Department of Medical Genetics, Institute of Health Science jointly-funded by Shanghai Institutes for Biological Sciences of Chinese Academy of Sciences and Shanghai Second Medical University, Shanghai 200025, China [3]State Key Laboratory of Medical Genomics, Rui-Jin Hospital Affiliated to Shanghai Second Medical University, Shanghai 200025, China

出　　处：《Chinese Science Bulletin》2006年第18期2231-2235,共5页

基　　金：the grants from Science ＆ Technology Committee of Shanghai Municipality （Grant No. 03DZ14022）; the National High Technology Research and Development Program of China （Grant No. 2001AA216081）; the National Natural Science Foundation of China for 0utstanding Young Scientists （Grant No. 39925023）.

摘　　要：Generation of transgenic mice by DNA microinjection has become one of the most important technologies in studying gene function in vivo. Ran- dom integration of transgene often results in inser- tional mutation which may complicate phenotype analysis of transgenic mice and/or create a good opportunity to study the function of endogenous gene. However,the utilization of these potentially valuable resources is hampered due to lack of efficient ap- proaches for rapid identification of multi-copy trans- gene integration sites. Here we propose two PCR-based approaches to identifying trans- gene/genome junction sequences. The efficiency of these two strategies was tested in 9 independent transgenic mouse lines. Among them,the transgene in 8 mouse lines was clearly localized to certain chromosome regions. These rapid and efficient ap- proaches will greatly facilitate the study of insertional mutation due to transgene integration.Generation of transgenic mice by DNA microinjection has become one of the most important technologies in studying gene function in vivo. Random integration of transgene often results in insertional mutation which may complicate phenotype analysis of transgenic mice and/or create a good opportunity to study the function of endogenous gene. However, the utilization of these potentially valuable resources is hampered due to lack of efficient approaches for rapid identification of multi-copy transgene integration sites. Here we propose two PCR-based approaches to identifying transgene/genome junction sequences. The efficiency of these two strategies was tested in 9 independent transgenic mouse lines. Among them, the transgene in 8 mouse lines was clearly localized to certain chromosome regions. These rapid and efficient approaches will greatly facilitate the study of insertional mutation due to transgene integration.

关 键 词：转基因 结合部位 插入突变 PCR 

分 类 号：Q78[生物学—分子生物学]