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. 2021 Mar 23;6(2):e00903-20. doi: 10.1128/mSystems.00903-20

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Copyright © 2021 Orozco-Hidalgo et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.

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FIG 3 — Supercharged SpyCatcher allows for secretion of synthetic extracellular matrix proteins. (A) Schematic of genome variants for secretion of SpyCatcher (SC)-ELP₆₀-336c protein and supercharged SpyCatcher [SC⁽⁻⁾]-ELP₆₀-336c protein. The green segment indicates a FLAG tag and orange segment indicates a Strep tag. All genes contain the native rsaA promoter and 5′ UTR. (B) Immunoblot with anti-FLAG antibodies against the whole-cell lysate (indicated by C) and extracellular medium (indicated by E) of Sec:parent (first and second lanes), Sec:SC-ELP₆₀-336c (third and fourth lanes), and Sec:SC⁽⁻⁾ELP₆₀-336c (fifth and sixth lanes). While the matrix protein is not detected in the Sec:parent and Sec:SC-ELP₆₀-336c cultures, the Sec:SC⁽⁻⁾-ELP60-336c strain achieves notable levels of secretion.