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. 2014 Jul;196(13):2333–2347. doi: 10.1128/JB.01572-14

FIG 11.

FIG 11

Secretion of AMA1, NCR, and MrVIA peptides fused to a His-tagged FlgM secretion signal. (A) Schematic diagram of the constructs tested. (B) Coomassie-stained SDS-PAGE gel of the supernatant from a culture expressing FlgM-His6-ETK-AMA1 in an optimized secretion strain (TH20685). The supernatant was concentrated 20-fold by TCA precipitation. (C) Western blot against 20 μl of nonconcentrated supernatant from panel B with anti-His antibody. (D) Coomassie-stained SDS-PAGE gel of the supernatant from cultures expressing three different NCR peptides fused to a His-tagged FlgM secretion signal in an optimized secretion strain background (NCR247, TH20221; NCR057, TH20222; NCR224, TH20223). The supernatants were concentrated 20-fold by TCA precipitation. (E) Coomassie-stained SDS-PAGE gel of the supernatant from a culture expressing conopeptide MrVIA fused to a His-tagged FlgM secretion signal in an optimized secretion strain background (TH20492). The supernatants were concentrated 20-fold by TCA precipitation. (F) Western blot against 20 μl of nonconcentrated supernatant from strain TH20492 (FlgM-His-ETK-MrVIA) with anti-His antibody grown under different arabinose induction concentrations.