Even though the end of iGEM 2011 is near, there is still a lot we would like to accomplish. By providing these toolkits, we hope to encourage future iGEM teams to experiment with Gibson cloning to help create large-scale constructs like the genome of Magnetotactic Bacteria.
Gibson Assembly Toolkit
1. Remove XhoI site from pGA3K3 vector:
2. Remove homologous regions from pGA4A5, pGA4C5 vectors: these homologous regions within the backbone are self-complementary and can bind to one another, circularizing the backbone without an insert. This could potentially reduce the backbones efficiency and should be eliminated.
3. Finally, we would like to continue adding more BioBrick compatible Gibson Vectors to help expand their popularity within the iGEM community.
There is still much work to be done before we can achieve our goal of making Magnetocoli; however, we feel our toolkit provides an essential foundation for future magnetosome gene manipulation.
1. Co-express the genes in E.coli to determine their gene interactions.
2. Build the scaffold required for proper magnetosome alignment.
- The construction of this scaffold would prove highly applicable to many other systems. For our "Make It" project concerning alkane production, we mentioned our interest in enzyme localization. Using mamI, a membrane localized gene, we hope to devise a system that would allow the localization of AAR and ADC to the fatty-acid rich membrane, thus increasing the efficiency of alkane production.
3. Continue to construct the full mamAB operon of the MAI in thirds.
4. Determine other regions of the MAI that contain additional essential genes for magnetosome formation.
Overall, the ability to produce and control uniform, nano-sized magnetic particles is attractive in areas such as medical imaging and nano-electronics where scientists and engineers are actively seeking innovative solutions for advancements in size and accuracy. Thus, if we are able to produce magnets in an organism that are thoroughly understood in a controlled manner, and be able to extract the magnets from them, this indeed is going to be very useful many projects in synthetic biology.