Fig. 1.

Identification of genes involved in OMV biogenesis. (A) Western blot against anti-polyHis showing total membrane (TM) and OMV fractions from BT WT constitutively expressing Inulinase fused to Nluc with a C-terminal 6xHis tag (INL-Nluc). This blot demonstrates that INL-Nluc is stably expressed and properly localizes to the OMV fraction. Samples were normalized by OD600 and ran on 10% SDS-PAGE. (B) Filtered supernatants from cells expressing INL-Nluc display higher luminescent output than those expressing Nluc in the cytoplasm. Filtered supernatants from overnight cultures (~20 h) were used for NanoGlow assays to quantify OMV production in vitro. Total luminescent output was normalized by OD600. (C) Schematic of OMV screening methodology (Created with https://BioRender.com).