Construction of Promoter-RBS Libraries and Its Application to Gene Circuit Design with Design Specifications

• Main Authors: Wu, Chih-Hung, 巫志鴻
• Other Authors: Chen, Bor-Sen
• Format: Others
• Language: en_US
• Published: 2012
• Online Access: http://ndltd.ncl.edu.tw/handle/21793418794487926123

博士 === 國立清華大學 === 電機工程學系 === 100 === A major goal of synthetic biology is to engineer synthetic gene circuits with desired behaviors. Up to now, however, it still requires extensive and iterative work. The main obstacle is the lack of some well-characterized biological parts and design methods. By the identification technique, the biological parts can be well-characterized to build a useful library for engineering a novel gene circuit. Hence how to build a useful library of biological parts for engineering synthetic gene circuit in vitro and in silico is an important topic from the viewpoint of synthetic biology. Promoters and RBSs are regarded as a lumped component in this study so that the promoter-RBS component can help us construct useful promoter-RBS libraries by the identification technique. The promoter-RBS libraries can be easily used to engineer a synthetic gene circuit before laborious process of trial-and-error. In general, there exist 3 kinds of promoter-RBS components in synthetic biology, hence 3 kinds of promoter-RBS libraries are constructed in this study, i.e., constitutive, repressor-regulated and activator-regulated promoter-RBS libraries.We provide the construction procedure for the constitutive, repressor-regulated, and activator-regulated promoter-RBS libraries. After building 9 promoter-RBS libraries, we provide the characteristic indexes of these libraries through their mathematical models, a systematic method is proposed to help us select the adequate promoter-RBS component set. Finally, we provide a library-based search method to help us quickly select the most adequate promoter-RBS component set from promoter-RBS libraries. The proposed library-based search method can reduce the number of trial-and-error experiments in selecting an adequate promoter-RBS component set for a synthetic gene circuit, and then a gene circuit can be predictably implemented by a systematic design method before trial-and-error experiments.