Synthetic Gene Involving Azobenzene-Tethered T7 Promoter for the Photocontrol of Gene Expression by Visible Light
Synthetic Gene Involving Azobenzene-Tethered T7 Promoter for the Photocontrol of Gene Expression by Visible Light
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DOI：
10.1021/sb5001092
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发表时间：
2015-04-01
影响因子：
4.7
通讯作者：
Asanuma, Hiroyuki
中科院分区：
文献类型：
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作者：
Kamiya, Yukiko;Takagi, Toshiki;Asanuma, Hiroyuki
在目前的研究中，我们展示了在一个无细胞翻译系统中，T7启动子在特定位置含有两个偶氮苯衍生物对基因表达的光调节。作为光开关，我们制备了偶氮苯-4‘-羧酸(AZO)和2，6-二甲基偶氮苯-4’-羧酸(DM-AZO)，它们在紫外光(波长370 nm)下从反式异构化为顺式异构化；4-甲硫基偶氮苯酸(S-偶氮酸)和2，6-dimethyl-4(methylthio)azobenzene-4‘-carobxylic酸(S-DM-偶氮酸)在4 0 0 nm可见光照射下顺式异构化。用S偶氮或S偶氮DM修饰的启动子可以用无害的可见光诱导绿色荧光蛋白的表达，而用偶氮或偶氮DM修饰的启动子只能用紫外光(340~360 nm)诱导绿色荧光蛋白的表达。因此，在无细胞翻译系统中实现了对绿色荧光蛋白生产的高效光调控，该系统具有可见光而不会造成光损伤。
In the present study, we demonstrate photoregulation of gene expression in a cell-free translation system from a T7 promoter containing two azobenzene derivatives at specific positions. As photoswitches, we prepared azobenzene-4'-carbcodyic acid (Azo) and 2,6-dimethylazobenzene-4'-carboxylic acid (DM-Azo), which were isomerized from trans to cis upon irradiation with UV light (lambda < 370 nm), and 4-methylthioazobenzene-4'-carboxylic acid (S-Azo) and 2,6-dimethyl-4(methylthio)azobenzene-4'-carobxylic acid (S-DM-Azo), which were cis-isomerized by irradiation with 400 nm visible light. Expression of green fluorescent protein from a promoter modified with S-Azo or S-DM-Azo could be induced by harmless visible light whereas that from a promoter modified with Azo or DM-Azo was induced only by UV light (340-360 nm). Thus, efficient photoregulation of green fluorescent protein production was achieved in a cell-free translation system with visible light without photodamage.