通过将工程学思想引入生物学，合成生物学旨在解决人类面临的健康、环保和能源等问题。新元件和新方法的开发与应用将推动合成生物学快速发展。申请人前期以线性双链DNA（PCR扩增产物）为元件在细胞及动物水平成功构建了与门逻辑基因线路。通过PCR反应将基因表达序列拆分成2~3个线性双链DNA分子以作为输入，新基因线路实现了低噪信比与门逻辑运算。在此基础上本课题拟开展以下研究：1）在不同细胞系及动物水平实现2输入、3输入与门逻辑运算，为建立一种基于线性双链DNA与门逻辑基因线路的合成生物学新方法提供依据；2）通过优化转染条件及双链DNA化学修饰提高与门运算输出信号，并在基因水平寻找可影响DNA连接效率的调控因子；3）分析DNA分子连接处的碱基插入缺失情况，利用线性双链DNA与门运算构建工程抗体库。本研究的完成将为合成生物学基因线路设计带来全新的元件与方法，同时为新蛋白生成提供新策略。

By introducing engineering principle into biology, synthetic biology aims to address the challenging issues, such as healthcare, environmental protection and energy. The development and application of novel elements and methods will promote the development of synthetic biology. Our preliminary results showed the successful construction of linear-double-stranded DNA (PCR amplicon) based AND logic calculation genetic circuits. Taking 2~3 different PCR amplicons (split from gene expression sequences) as inputs, our novel genetic circuit design principle achieves low noise-signal ratio AND gate calculation. Basing on these preliminary results, current project will focus on the following aspects: 1) to construct 2-input or 3-input AND logic genetic circuits in vitro and in vivo; to establish a novel synthetic biology tool for this project. 2) to improve the AND gate output signal through transfection condition optimization and double-stranded DNA chemical modifications; to explore genes involved in this AND gate calculation. 3) to analyze nucleotide(s) insertion or deletion at the junction between double-stranded DNA molecules; to construct linear-double-stranded DNA based AND gate calculation-based engineering antibody library. This project will bring novel elements and methods to synthetic genetic circuits design and provide novel strategies for new protein genesis.