System design of artificial genetic circuit by integration of dry and wet experiment

干湿实验一体化人工遗传电路系统设计

• 批准号: 20300102
• 负责人: HANAI Taizo
• 金额: $ 12.31万
• 依托单位: Kyushu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2008
• 资助国家: 日本
• 起止时间: 2008 至 2010
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-20300102/
• 关键词: 生体生命情報学; 生物・生体工学; システム生物学; 合成生物学; システム解析

By using lambda phage promoters, construction of oscillatory genetic circuit was carried out. From the preliminary experiment, the expressional weakness of pRM promoter is problem for the oscillation. In order to increase the expression level from pRM promoter, several mutations of pRM and RBS with higher translation efficiency were applied to this circuit. Despite these efforts, we could not get the continuous oscillation.By using lac and tet repressors, we tried to construct a toggle switch. placIq-lacI gene was introduced into genome, on low copy plasmid, or medium copy plasmid. The toggle switch on high copy plasmid was transformed with each pLlacIq-lacI gene. As the result of the GFP expression experiment, only the strain with low copy plasmid as a source of lac repressor worked as we desired. Mathematical model was also constructed.

利用λ噬菌体启动子构建振荡遗传回路。从初步实验来看，pRM启动子的表达弱是引起振荡的一个问题。为了提高pRM启动子的表达水平，我们将几种翻译效率较高的pRM和RBS突变应用到该电路中。尽管做了这些努力，我们还是没有得到连续的振荡。利用lac和tet阻遏因子，我们尝试构建一个拨动开关。将placIq-lacI基因分别导入基因组、低拷贝质粒和中拷贝质粒。用pLlacIq-lacI基因转染高拷贝质粒上的开关。在GFP表达实验中，只有以低拷贝质粒作为lac抑制源的菌株才能达到我们所期望的效果。建立了数学模型。

项目成果

遺伝子トグルスイッチの試作とその特性

基因拨动开关的样机制作及其特点

• 发表时间: 2010
• 作者: N. Shimizu;M. Hagiwara;Y. Ogawa;K. Toyama;H. Nakagawa;花井泰三・安武俊輔・岡本正宏
• 通讯作者: 花井泰三・安武俊輔・岡本正宏

遺伝子トグルスイソチの構築と解析

基因切换isochi的构建与分析

• 发表时间: 2010
• 作者: Y. Ogawa;K. Imai;K. Toyama;安武俊輔・花井泰三
• 通讯作者: 安武俊輔・花井泰三