We introduced a self-assembly synthetic adhesion system by transfecting initimin-Nb2 fusion into ''E. coli''. Initimin-Nb2 fusion is composed of a surface display system(intimin) and the coding sequence of a nanobody. The surface display system, which includes a short N-terminal signal peptide to direct its trafficking to the periplasm, a LysM domain for peptidoglycan binding, and a beta-barrel for transmembrane insertion<ref>Piñero-Lambea, C., Bodelón, G., Fernández-Periáñez, R., Cuesta, A. M., Álvarez-Vallina, L., & Fernández, L. Á. (2015). Programming controlled adhesion of E. coli to target surfaces, cells, and tumors with synthetic adhesins.'' ACS Synthetic Biology, 4''(4), 463–473. https://doi.org/10.1021/sb500252a </ref>, possesses the outer membrane anchoring of the nanobody<ref>Glass, D. S., & Riedel-Kruse, I. H. (2018). A Synthetic Bacterial Cell-Cell Adhesion Toolbox for Programming Multicellular Morphologies and Patterns. ''Cell, 174''(3), 649-658.e16. https://doi.org/10.1016/j.cell.2018.06.041</ref>.
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We’ve constructed this fusion protein into our ribozyme-assisted polycistronic co-expression system：pRAP.
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===Usage and Biology===
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Similar to [https://parts.igem.org/Part:BBa_K4765106 BBa_K4765106], we replaced Nb3 with Nb2 in order to interact with [https://parts.igem.org/Part:BBa_K4765103 BBa_K4765103] to create a biofilm.
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The surface-displayed nanobody can specifically interact with the antigen produced by [https://parts.igem.org/Part:BBa_K4765103 ribozyme+strong RBS+intimin-Ag2 fusion+stem-loop] .In our project, we took full advantage of the Ag-Nb interaction to create a biofilm with a programmable physical structure<ref>Kim, H., Skinner, D. J., Glass, D. S., Hamby, A. E., Stuart, B. A. R., Dunkel, J., & Riedel-Kruse, I. H. (2022). 4-bit adhesion logic enables universal multicellular interface patterning. ''Nature, 608''(7922), 324–329. https://doi.org/10.1038/s41586-022-04944-2</ref>.
