Methods

Here you can find a list of methods for working with Bacillus subtilis:

Analysis of protein-protein interactions

• Bacterial Two Hybrid: BACTH

• SPINE: A method to detect in vivo protein-protein interactions

• in vivo detection of protein-protein interactions using DivIVA and GFP PubMed

Analysis of membrane proteins

• application of a split green fluorescent protein reporter PubMed

Protein expression

• the LIKE system PubMed

Strain construction

• introduction of markerfree deletions: PubMed

• generation of markerfree mutations: PubMed and PubMed

RNA analysis

• Mechanical cell disruption and extraction of RNA from B. subtilis: Media:SOP-RNA.pdf

Genetic work with non-standard strains

• transformation of wild-type B. subtilis strains incl. NCIB3610: PubMed
• efficient electroporation of B. subtilis PubMed
• a novel transformation protocol for B. subtilis DB104 PubMed

General procedures

• Standard operation procedures (SOPs) of the SYSMO-BACELL consortium

Live cell imaging

• Live cell imaging of B. subtilis cells using automated time-lapse microscopy PubMed
• Assay of gene expression dynamics using live cell imaging PubMed

Transposon mutagenesis

• mariner transposon mutagenesis for random inducible-expression insertions and transcriptional reporter fusion insertions PubMed

Protein-DNA interactions

• ChAP-chip: A modified ChIP-chip protocol for the in vivo identification of binding sites of DNA-binding proteins PubMed

Key references

Additional publications: PubMed

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Arturo Rodríguez-Banqueri, Lukasz Kowalczyk, Manuel Palacín, José Luis Vázquez-Ibar
Assessment of membrane protein expression and stability using a split green fluorescent protein reporter.
Anal Biochem: 2012, 423(1);7-14
[PubMed:22285978] [WorldCat.org] [DOI] (I p)