We have finished constructing all the parts(BBa_K2684000- BBa_K2684006) which have all been sequenced and submitted to the iGEM parts registry
|BBa_K52684000||CotA laccase of B. subtilis||CotA of B. subtilis is a copper-dependet laccase, which can catalyze the oxidation of board range of synthetic dyes.|
|BBa_K52684001||PelB-CotA||PelB signal peptide linking with CotA to transfer CotA out of the bacteria.|
|BBa_K52684002||PhoA-CotA||PhoA signal peptide linking with CotA to transfer CotA out of the bacteria.|
|BBa_K52684003||OmpA-CotA||OmpA signal peptide linking with CotA to transfer CotA out of the bacteria.|
|BBa_K52684004||SpyCatcher-sfGFP||SfGFP fused to the SpyCatcher domain can be covalently attached onto the biofilm displaying SpyTag.This part is used to test our immobilization strategy of CotA.|
|BBa_K52684005||SpyCatcher-CotA||CotA fused to the SpyCatcher domain can be covalently attached onto the biofilm displaying SpyTag.|
|BBa_K52684006||CsgA-SpyTag||CsgA is a major subunit of the biofilm of E. coli(1)(2). A SpyTag is fused to CsgA so that CotA laccase can be fixed on biofilm through SpyTag-SpyCatcher chemistry.|
The favorite part of pur team is BBa_K2684006. We improved it from previous part BBa_K1583000. CsaA is the main component to form biofilm, and by adding SpyTag which was fused onto the csgA sequence, we could fix the CotA laccase with SpyCatcher. Covalent bond would be formed between SpyTag and SpyCatcher.
a. Improve Previous Parts
(1). Wang, X, et al. “Programming Cells for Dynamic Assembly of Inorganic Nano-Objects with Spatiotemporal Control.” Advanced Materials (Deerfield Beach, Fla.)., U.S. National Library of Medicine, Apr. 2018, www.ncbi.nlm.nih.gov/pubmed/29516606.
(2). Chen, Allen Y., et al. “Synthesis and Patterning of Tunable Multiscale Materials with Engineered Cells .” Nature Materials, U.S. National Library of Medicine, May 2014, www.ncbi.nlm.nih.gov/pmc/articles/PMC4063449/.