Project description:The aim of the experiments was to determine the regulon of the Bacillus subtilis alternative sigma factor SigI. Biological relevance: To expand our knowledge about Bacillus subtilis transcriptional network under unfavorable conditions. Experimental workflow overview: Bacillus subtilis 168 trp+ (BaSysBio) was used as the genetic background. (i) sigI-rsgI knock-out, (ii) rsgI knock-out, and (iii) wr strains were cultured in LB medium to mid-exponential phase at 37°C and 52°C. Total RNA was isolated from 3 ml of the culture. rRNA was depleted from the samples with RiboMinus; subsequently RNA-seq libraries were prepared (Illumina compatible NEXTflex Rapid Directional RNA-Seq Kit, Bioo Scientific) and sequenced at the EMBL GeneCore facility. The experiment was performed in three independent replicates.
Project description:Alanine dehydrogenase (OF4Ald) from the alkaliphilic Bacillus pseudofirmus OF4 was expressed and purified with a His6 tag in a form suitable for X-ray crystallographic analysis. Crystals were grown by the hanging-drop vapour-diffusion method at 289?K using a solution consisting of 0.1?M Tris-HCl pH 8.0, 0.2?M LiSO4, 22%(w/v) PEG 3350. X-ray diffraction data were collected to 2.8?Å resolution. The crystal belonged to the triclinic space group P1, with unit-cell parameters a = 88.04, b = 105.59, c = 120.53?Å, ? = 88.37, ? = 78.77, ? = 82.65°.
