Team:Bielefeld-CeBiTec/Parts
Parts Overview
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For the assembly of our Troygenics, we are using a two plasmid system. To keep both plasmids inside of the cell, we developed a novel chloramphenicol split resistance. Since chloramphenicol is the standard antibiotic used by the iGEM community, this part will be beneficial for all future iGEM teams.
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To build the Troygenics, we designed many composite parts for different sections of our project. Our best composite part was the completed Troygenic with an endocytosis ligand as a lab application containing GFP.
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We have improved the function of chloramphenicol acetyltransferase by modifying it for it to be useful in a broader spectrum of applications. By choosing a split site for the protein and fusing each part with Npu DnaE Intein, it is possible to split an antibiotic resistance gene and use it for a two-plasmid-based system. Thereby, instead of working with many antibiotics, only one is needed. This is an innovative way to overcome the problem of multi resistant bacteria.
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Would you love working with a two-plasmid system using only one antibiotic resistance? We enable you to do this by splitting antibiotic resistance genes with intein-mediated protein trans-splicing. By implementing this innovative approach, the likelihood of spreading resistances is decreased, as one plasmid does not encode the full protein. To ensure the universal applicability within the iGEM community, we selected resistances against commonly used antibiotics: Chloramphenicol, Kanamycin, Ampicillin, and Hygromycin B. The optimal split point was selected based on our model. By constructing and characterizing a collection of two-plasmid systems we demonstrated the feasibility of our split-antibiotic system to maintain two plasmids in a single cell.
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A very common method of visualizing proteins in vivo and in vitro are fluorescence reporters. We have improved the characterization of the commonly used red fluorescent protein mCherry in the iGEM parts registry.
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