Team:SHSBNU China/Hardware

Parts

Parts

I. Overview

We have finished constructing all the parts(BBa_K2684000- BBa_K2684006) which have all been sequenced and submitted to the iGEM parts registry

Name Description Function
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.

a. Improve Previous Parts

We improved partBBa_K1583000 by adding a sequence of SpyTag in order to fix CotA laccase onto the biofilm.

We improved partBBa_K1336002 by eliminating its EcoRI cutting site.

II. References

(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/.