Construction of a genetic AND gate under a new standard for assembly of genetic parts.
Construction of a genetic AND gate under a new standard for assembly of genetic parts.
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DOI：
10.1186/1471-2164-11-s4-s16
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发表时间：
2010-12-02
期刊：
影响因子：
4.4
通讯作者：
Kiga D
中科院分区：
文献类型：
--
作者：
Ayukawa S;Kobayashi A;Nakashima Y;Takagi H;Hamada S;Uchiyama M;Yugi K;Murata S;Sakakibara Y;Hagiya M;Yamamura M;Kiga D
适当调节各个基因的表达是在活细胞内实现人工生物系统的瓶颈。需要修饰几个启动子序列以实现系统的适当调节。然而，将操纵子（用于基因表达的蛋白质的结合位点）插入质粒的基因调控区需要耗时的过程。因此，需要将操纵子序列整合到质粒的调控区的标准化方法。我们开发了一种标准化的方法，用于整合操纵基因序列到质粒的调控区，并构建了一个合成的启动子，作为遗传与门。通过对质粒调控区和操纵子的标准化，建立了操纵子模块化组装平台。此外，通过在调控区上组装两个不同的操纵子部分，我们构建了具有与门功能的调控装置。我们实施了一个新的标准来组装用于构建功能遗传逻辑门的操作符部件。分子尺度上的逻辑门对细胞行为的重编程具有重要意义。
Appropriate regulation of respective gene expressions is a bottleneck for the realization of artificial biological systems inside living cells. The modification of several promoter sequences is required to achieve appropriate regulation of the systems. However, a time-consuming process is required for the insertion of an operator, a binding site of a protein for gene expression, to the gene regulatory region of a plasmid. Thus, a standardized method for integrating operator sequences to the regulatory region of a plasmid is required. We developed a standardized method for integrating operator sequences to the regulatory region of a plasmid and constructed a synthetic promoter that functions as a genetic AND gate. By standardizing the regulatory region of a plasmid and the operator parts, we established a platform for modular assembly of the operator parts. Moreover, by assembling two different operator parts on the regulatory region, we constructed a regulatory device with an AND gate function. We implemented a new standard to assemble operator parts for construction of functional genetic logic gates. The logic gates at the molecular scale have important implications for reprogramming cellular behavior.