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

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

Work with non-domesticated strains

• transformation of wild-type B. subtilis strains incl. NCIB3610: 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

Jonathan W Young, James C W Locke, Alphan Altinok, Nitzan Rosenfeld, Tigran Bacarian, Peter S Swain, Eric Mjolsness, Michael B Elowitz
Measuring single-cell gene expression dynamics in bacteria using fluorescence time-lapse microscopy.
Nat Protoc: 2011, 7(1);80-8
[PubMed:22179594] [WorldCat.org] [DOI] (I e)

Eric R Pozsgai, Kris M Blair, Daniel B Kearns
Modified mariner transposons for random inducible-expression insertions and transcriptional reporter fusion insertions in Bacillus subtilis.
Appl Environ Microbiol: 2012, 78(3);778-85
[PubMed:22113911] [WorldCat.org] [DOI] (I p)

Imke G de Jong, Katrin Beilharz, Oscar P Kuipers, Jan-Willem Veening
Live Cell Imaging of Bacillus subtilis and Streptococcus pneumoniae using Automated Time-lapse Microscopy.
J Vis Exp: 2011, (53);
[PubMed:21841760] [WorldCat.org] [DOI] (I e)

Christina Herzberg, Lope Andrés Flórez Weidinger, Bastian Dörrbecker, Sebastian Hübner, Jörg Stülke, Fabian M Commichau
SPINE: a method for the rapid detection and analysis of protein-protein interactions in vivo.
Proteomics: 2007, 7(22);4032-5
[PubMed:17994626] [WorldCat.org] [DOI] (P p)

Shu Ishikawa, Yoshitoshi Ogura, Mika Yoshimura, Hajime Okumura, Eunha Cho, Yoshikazu Kawai, Ken Kurokawa, Taku Oshima, Naotake Ogasawara
Distribution of stable DnaA-binding sites on the Bacillus subtilis genome detected using a modified ChIP-chip method.
DNA Res: 2007, 14(4);155-68
[PubMed:17932079] [WorldCat.org] [DOI] (P p)