啟動子-核糖體結合位庫的建立與基因電路規格設計上的應用 於藍綠菌細胞 PCC 7942

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

李佳倫(Li, Jia Lun)

指導教授 ： 陳博現

國立清華大學/電機資訊學院/電機工程學系/碩士(2015年)

摘要

至今在藍綠菌細胞中為了能夠系統化得設計基因電路，建立啟動子-核糖體的元件庫，以因應各種廣泛的應用仍是一件吸引人的事。在藍綠菌細胞 PCC 7942 中，為了能夠系統化得設計基因電路，我們建立了三種啟動子-核糖體結合位元件庫，即連續表現型、抑制子調控型及促進子調控型。將工程化基因電路轉殖到藍綠菌中，分別提供了這三種啟動子-核糖體結合位庫的建立流程圖，根據實驗中量測到啟動子-核糖體的強度和動態模型，啟動子-核糖體被視為一種特性組件，再來透過系統鑑別參數的技術將生物元件特性化，並且調控不同種類的生物組件，用於基因電路設計以符合在藍綠菌細胞中的期望行為。在藍綠菌細胞中，展現了設計的示範例子關於細胞族控制，說明了設計流程根據所提出的啟動子-核糖體搜尋方法，並且結合活體細胞做驗證。為了使設計更加方便，利用系統化方法來控制從元件庫中選擇出的核糖體元件強度，以滿足在藍綠菌細胞中的行為。因此我們相信所提出系統化的工程設計方法，能提供快速發展的合成生物領域，讓使用者能選擇最佳的元件以實現各種的應用。

關鍵字

藍綠菌 ； 啟動子-核糖體結合位庫 ； 細胞族群控制

並列摘要

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.

並列關鍵字

cyanobacteria ； promoter-RBS library ； cell population control

參考文獻

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