Construction of Promoter-RBS Libraries of the Cyanobacterium Synechococcus sp. PCC 7942 and Their Application to Systematic Synthetic Circuit Design to Match User-Oriented Specifications

• Main Authors: Li, Jia Lun, 李佳倫
• Other Authors: Chen, Bor Sen
• Format: Others
• Language: en_US
• Published: 2015
• Online Access: http://ndltd.ncl.edu.tw/handle/56822793541989417529

碩士 === 國立清華大學 === 電機工程學系 === 103 === At present, it is still appealing to have promoter-RBS libraries for systematic design of synthetic genetic circuit in Cyanobacteria for a variety of application. In the Cyanobacterium Synechococcus sp. strain PCC 7942, we set up three kinds of promoter-RBS libraries, i.e., constitutive, repressor-regulated and activator-regulated promoter-RBS libraries, for systemic design of synthetic gene circuits in Cyanobacteria. In order to engineer synthetic gene circuit into Cyanobacterium Synechococcus sp. PCC 7942, we provide the flow chart to constructing constitutive, repressor-regulated, and activator-regulated promoter-RBS libraries based on promoter-RBS intensities through experimental data, i.e., , , and , respectively. By the parameter identification technique based on dynamic model, a promoters-RBS is regarded as a characteristic part to be selected for gene circuit design with a desired behavior in Cyanobacteria, and regulated by different biological parts in blue-green algae. A design example with desired cell population control is given to illustrate the design procedure of synthetic circuit in Cyanobacteria by the proposed promoter-RBS library-search method and is validated by experimental data in vivo. For the convenience of design, we provide a systematic method for a genetic circuit to control part’s intensity to achieve some desired behavior in Cyanobacterium Synechococcus sp. PCC 7942. Consequently, we believe that the proposed library-based search method can help user select an adequate set of promoter-RBS parts to realize various kinds of application in rapidly developed synthetic biology.