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Team:Alberta
From 2009.igem.org
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BioBytesAt present, the cost to synthesize oligonucleotides has been continually declining and therefore their availability is exponentially growing. However, the current technique to ligate pieces of DNA together is outdated. Present methods take a considerable amount of time to piece DNA together, making large constructs incredibly difficult to build. The University of Alberta 2009 iGEM team would like to introduce the BioBytes Chromosome Assembly System. This method refers to a mechanism for rapid and reliable construction of plasmids (i.e. artificial gene sets) in vitro. It allows for the assembly of components in a structured manner within hours rather than days. Our hope is to see this method become a valuable tool for any molecular biologist. Furthermore we have adapted our approach for Biofabrication by developing a robot which can use our method for automated assembly. Finally, microfluidics have been utilized to miniaturize construction allowing for additional validation for automated production of constructs. The method can be applied to numerous different applications, however, its greatest application is for the assembling of entire genomes. For this reason we have provided a detailed explanation regarding the requirements of constructing a minimal genome including an in-silico method for identifying essential genes in any organism, and a theoretical design of replacing the host chromosome with the new synthetic genome. |
The Minimal Genome ProjectThe minimal Escherichia coli genome has been the holy grail of biology for a number of years. E. coli is the most widely used cellular research tool by the molecular biology community. Since scientific research is based upon reductionism and simplification for understanding, a simplified version of an experimental model organism such as E. coli is ideally preferred as a chassis for experimentation. Reducing the E. coli genome to roughly 10% of its original size, demonstrates a great simplification of this model organism. To reconstruct such an organism, we plan on building an artificial E. coli chromosome using the BioBytes chromosome assembly system and inserting it into living E. coli cells. We then intend to remove the host chromosome by making it incapable of division, thus allowing only the artificially inserted chromosome to propagate through multiple generations as the cells grow and divide. This is markedly different from the current, time-consuming method of knocking out inessential genes, one at a time, in an effort to produce the minimal genome. It is this difference that we hope to exploit in our attempt to win the race to produce the minimal E. coli genome. |
Team AchievementsThrough our efforts we have made the following accomplishments:
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