Team:BHSF ND/Parts
2019 IGEM BHSF_NDParts
Overview
We submitted 68 parts, with 40 composite parts, and 28 basic parts. Don’t think we win at quantity but we actually provided a series of data with high-quality. In our bistable experiment, our favorite new basic parts is BBa_K3202022 which is a green fluorescence protein to demonstrate the function of our bistable system. And our favorite composite parts is BBa_K3202045 and it is designed to achieve zero basal expression in the absence of any inducer while ensuring constant transcriptional capacities and induction rates among promoters. We also included 28 part in our best part collection. Check the BHSF_ND parts family.
Parts Collection
In our study, we designed a digitalizing bistable module that can enable conditional suicide of engineered bacteria in factories to ensure the protection of intellectual properties. Therefore, we spliced our work into four different modules: an leakage/inducer module, a recombinase-based flipping module, a toxin module, and a bistable module. To investigate and improve the four modules, we build various libraries to support our study. For the recombinase module, we created composite parts BBa_K3202032-BBa_K3202043 (except for BBa_K3202042) to examine which of the three recombinases can produce the best results and to improve the RBS of the recombinases and thereby promote their functions. Also, we optimized the toxin module, for which we built BBa_K3202052-BBa_K3202054 and BBa_K3202066-BBa_K3202068 so that a variety of tool pack for killing the different bacteria is created, and their compatibility with the bistable system is measured. Likewise, other parts included in this part collection are used to improve the other two modules. Additionally, We tested and compared two brand new RBS for iGEM parts (RBS that form scaffold formation) and a common one in our bistable system to ensure there is no leakage. Overall, our team contributed 68 parts to the iGEM parts library and quantitatively tested and optimized the parts we contributed.
Basic Parts
We introduced this new part to the iGEM parts, and have characterized this part in several of our composite parts. This part is used to demonstrate the function of our bistable system. The newly introduced msfGFP is very sensitive according to our results. Comparing to BN006/Contains BBa_K3202029/Backbone, this new part shows significant difference in both AraC-pBAD inducer system and Xyls-Pm inducer system. Also, comparing to sfGFP, this new part shows significant difference in both AraC-pBAD inducer system and Xyls-Pm inducer system as well. We provided qualitative data and quantitative data in the part registry page.
Composite Parts
The complete sequence of this composite part is Xyls-Pc-Pm-TetR-RBS1-msfGFP-T500-T1/TE-MicCsRNA-sRNA Binding Site-PLtetO. This composite part is designed as one of several sets of our bistable system, which is designed to achieve zero basal expression in the absence of any inducer while ensuring constant transcriptional capacities and induction rates among promoters. Compared with control groups and have its function measured on a different plasmid(other than the one it was originally on), we demonstrated that this system is a valid BioBrick and can be used in future synthetic biological studies.We provided qualitative data and quantitative data in the part registry page.