Parallel DNA assembly by recombination

© 2011 Dr. Zhenyu ShiThe development of synthetic biology requires the construction of much more sophisticated genetic systems; however, mathematical models are too simplified to produce accurate predictions. Therefore, successful constructions have to rely on trial and error. In order to simplify the constructions process and develop cost-efficient and convenient technology for batch construction tests, a new efficient parallel DNA assembly technology was established after testing several strategies. Recombination was chosen as a robust attachment assembly system as it does not require extreme conditions and can ensure efficient single copy integration and extraction. An experimental demonstration of assembly of five DNA fragments into a host genome was performed. A software platform for design and management of molecular cloning experiments was developed to manage construction of the assembly system. Promising technologies that are useful for recombination-based DNA assembly were evaluated. A DNA assembly theory was set up from the study of different strategies and provides a basis for design of DNA assembly systems and a guideline for future studies. This new family of methods has the potential advantages over all other methods, such as less or no in vitro operations, lower costs and shorter turnover period. This study is the first one to set up the theory and to attempt to extract the full potential of this technology based on the theory