Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
10532552 | Analytical Biochemistry | 2013 | 5 Pages |
Abstract
The enzyme ÏC31 integrase from Streptomyces phage has been documented as functional in mammalian cells and, therefore, has the potential to be a powerful gene manipulation tool. However, the activity of this enzyme is cell-type dependent. The more active mutant forms of ÏC31 integrase are required. Therefore, a rapid and effective method should be developed to detect the intracellular activity of ÏC31 integrase. We devised in this study an integrase-inversion cassette that contains the enhanced green fluorescent protein (EGFP) gene and the reverse complementary DsRed gene, which are flanked by attB and reverse complementary attP. This cassette can be inverted by ÏC31 integrase, thereby altering the fluorescent protein expression. Thus, ÏC31 integrase activity can be qualitatively or quantitatively evaluated based on the detected fluorescence. Furthermore, this cassette-based method was applied to several cell types, demonstrating that it is an efficient and reliable tool for measuring ÏC31 integrase activity in mammalian cells.
Related Topics
Physical Sciences and Engineering
Chemistry
Analytical Chemistry
Authors
Taian Liu, Yongxiang Fang, Huaijie Jia, Guohua Chen, Qisai Guan, Xiaobing He, Wenjuan Yao, Shuang Zeng, Zhizhong Jing,