Preparation of Bacillus subtilis cell samples and generation of an SDS-PAGE.
Thu Tm DaoDat D TruongLan Nh DuongNhi Ny NguyenHoang Duc NguyenPublished in: BioTechniques (2023)
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 OD 600 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, P grac212 . 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 .
Keyphrases
- bacillus subtilis
- randomized controlled trial
- induced apoptosis
- poor prognosis
- molecularly imprinted
- dna methylation
- single cell
- gene expression
- binding protein
- transcription factor
- crispr cas
- cell proliferation
- signaling pathway
- risk assessment
- long non coding rna
- oxidative stress
- endoplasmic reticulum stress
- stem cells
- mesenchymal stem cells
- bone marrow