Project description:We investigated the gene expression and metabolic regulatory mechanisms associated with the high-level accumulation of ICPs by performing the transcriptomics analysis of B. thuringiensis strain CT-43, using Illumina high throughout sequencing (RNA-seq) technique.
Project description:We investigated the gene expression and metabolic regulatory mechanisms associated with the high-level accumulation of ICPs by performing the transcriptomics analysis of B. thuringiensis strain CT-43, using Illumina high throughout sequencing (RNA-seq) technique. The bacterial cells were collected at the time points of 7 h, 9 h, 13 h and 22 h for the whole-genome transcriptomics, respectively.
Project description:To determine Sigma 54 (SigL) reglons in Bacillus thuringiensis HD73 strain, A sigLmutant, HD(ΔsigL::kan), was constructed with insertion of kanamycin resistance gene cassete. We have employed whole genome microarray expression profiling as a discovery platform to identify the difference of gene expression between mutant and wild-type strains.
Project description:In Bacillus thuringiensis CT-43, five insecticidal crystal proteins (ICPs, Cry protein) are encoded. We extracted the Cry proteins, ran the SDS PAGE (two Cry protein bands were observed), and tried to identify the composition of the two Cry protein bands in the SDS PAGE. The bioinformatics pipeline is described as follows: First, we converted the original mass spectrum files to the mgf file (peaks file), then the mgf files were searched against the Bacillusthuringiensis CT-43 protein database using Mascot (version 2.3.02). The search parameters were: i) trypsin was chosen as the enzyme with one missed cleavage allowed; ii) the fixed modifications of carbamidomethylation were set as Cys, and variable modifications of oxidation as Met; iii) peptide tolerance was set as 0.05 Da, and MS/MS tolerance was set as 0.1 Da. The peptide charge was set as Mr, and monoisotopic mass was chosen. An automatic decoy database search strategy was employed to estimate the false discovery rate (FDR). The FDR was calculated as the false positive matches divided by the total matches. In the final search results, the FDR was less than 1.5%.
Project description:To determine Sigma 54 (SigL) reglons in Bacillus thuringiensis HD73 strain, A sigLmutant, HD(M-NM-^TsigL::kan), was constructed with insertion of kanamycin resistance gene cassete. We have employed whole genome microarray expression profiling as a discovery platform to identify the difference of gene expression between mutant and wild-type strains. 2 ml samples were separately harvested from B. thuringiensis HD73 and HD(M-NM-^TsigL::kan) strains grown in SchaefferM-bM-^@M-^Ys sporulation medium (SSM) at stages T7 of stationary phase (7 hours after the end of the exponential phase). Three independent repeats were performed for each stain.
Project description:Transcriptional profiling of C. elegans nasp-1 / btr-1 mutant worms versus wild type N2 strain, both exposed to the bacterial pathogen Bacillus thuringiensis DB27.