Preparation of Bacillus subtilis cell samples and generation of an SDS-PAGE

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Abstract

Bacillus subtilis is a critical host for producing recombinant proteins. However, the SDS-PAGE process, including the sample preparation steps, varies among B. subtilis-related studies, making it impossible to compare findings. Hence, this paper provides a simple guide to culture and collect B. subtilis cells through an OD600 measurement and a protocol for SDS-PAGE. These techniques were applied to check the expression of a BgaB, a reporter protein and LukF-PV, a potential vaccine candidate against S. aureus, in the cytoplasm of B. subtilis under the control of a strong promoter, Pgrac212. This protocol could be helpful for scientists in preparing samples and generating an SDS-PAGE experiment, as well as favoring the unification of research about protein expression in B. subtilis.

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Dao, T. T. M., Truong, D. D., Duong, L. N. H., Nguyen, N. N. Y., & Nguyen, H. D. (2023). Preparation of Bacillus subtilis cell samples and generation of an SDS-PAGE. BioTechniques, 74(3), 123–129. https://doi.org/10.2144/btn-2022-0083

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