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Fig. 3 | Microbial Cell Factories

Fig. 3

From: Stable expression of HIV-1 MPER extended epitope on the surface of the recombinant probiotic bacteria Escherichia Coli Nissle 1917 using CRISPR/Cas9

Fig. 3

Whole cell dot bot and Western blot assay screening of surface expression of the MPER. (A) Dots representing whole cells immunostained with 2F5 primary antibodies of viable (V) or heat-killed (HK) EcN-MPER and EcN from passages 1 (P1) and 30 (P30) of the overnight cultures. The recombinant HIV-1 gp41 protein at concentrations of 15 and 150 ng was used as a positive control (PC). Detection was performed using the goat anti-human IgG IRDye® 800CW secondary antibody. (B) Western blot analysis of the outer membrane vesicles (OMVs) – rich fraction extracted from EcN-MPER culture supernatant. MPER was detected in the OMVs-rich fraction and positive control (EcN-MPER pellet) using HIV-1 gp41 (2F5) monoclonal antibody and rabbit anti-human HRP-conjugated IgG (Fc specific) secondary antibody. (C) DLS analysis of the OMVs fraction, and (D) negative stain TEM analysis of the same fraction

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Microbial Cell Factories

ISSN: 1475-2859

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