To help other teams to create their own surface-displaying bacteria, we have created and submitted several parts to the iGEM Registry. Check them out here!
Gold Medal: Best Composite Part
Caulobacter crescentus Parts
Our team contributed the following novel parts as was used in our C. crescentus workflow, check them out below. Further details of each part and the role it played in meduCA can be found on our page!
Our team thoroughly characterized the new protein coding composite part, BBa_25EMPHCH, which encodes for our most successful surface display fusion protein, PrsaA constitutive promoter+RsaA-BtCAI. This fusion protein will be anchored to the membrane of Caulobacter crescentus CB2A JS4038, enabling conversion of carbon dioxide into bicarbonate within the meduCA pipeline. All protocols designed and adapted by our team have also been made available on our page to allow for future use by other teams.
The fusion of RsaA with Bovine carbonic anhydrase II (BtCAII), codon optimized for E. coli. This part is expressed under the PrsaA constitutive promoter, and will be anchored to the membrane of Caulobacter crescentus CB2A, enabling conversion of carbon dioxide into bicarbonate.
The RsaA surface display protein from Caulobacter crescentus with Golden Gate compatible multiple cloning site for display of heterologous proteins.
Bronze Medal: Contributions
Synechococcus elongatus Parts
Our team contributed the following novel parts as was used in our S. elongatus workflow, check them out below. Further details of each part and the role it played in meduCA can be found on our page!
Terminator from clpP-clpX operon in E. coli. According to Gale et al., 2021, is strong terminator across E. coli, Synechococcus elongatus, and Synechocystis.
Short homology arm upstream of Synechococcus elongatus S-layer protein VCBS. When placed upstream of VCBS N domain in a construct for homologous recombination, combined length is sufficiently long for efficient recombination (~800 bp).
Synechococcus elongatus S-layer display suicide vector. Integrates fusion protein and kanamycin resistance gene into genome at locus of wild-type S-layer protein via homologous recombination. Has inducible counterselective marker using theophylline and IPTG. Insert CDS of any protein minus its stop codon using SapI Golden Gate Assembly. Has sfGFP reporter at empty state to verify insertion.
Synechococcus elongatus S-layer display shuttle vector based on pANS plasmid, confers kanamycin resistance. Insert CDS of any protein minus its stop codon using SapI Golden Gate Assembly. Has sfGFP reporter at empty state to verify insertion.
