Project description:The alkaliphilic halotolerant bacterium Bacillus sp. N16-5 often faces salt stress in its natural habitats. One-color microarrays was used to investigate transcriptome expression profiles of Bacillus sp. N16-5 adaptation reactions to prolonged grown at different salinities (0%, 2%, 8% and 15% NaCl) and the initial reaction to suddenly alter salinity from 0% to 8% NaCl.
Project description:RNAseq analysis was performed to evaluate gene expression differences between strains 1-9 and PAK-AR2.P. aeruginosa PAK-AR2 and 1-9 cells were grown to OD600 of 0.8 before harvesting. The collected cells were treated with RNAprotect Bacteria Reagent (Qiagen) and subjected to snap freezing in liquid nitrogen and delivered to BGI in dry ice for transcriptome resequencing analysis.The differentially expressed genes (DEGs) were determined between PAK-AR2 and 1-9 with the standards of false discovery rate (FDR ) ≤ 0.001, fold change |log2Ratio|≥1.A total of 4,355,305 reads matched to the referenced genome in the sample of PAK-AR2, and 3,544,484 reads in the sample of 1-9.Transcriptome data showed that expression of 361 genes were upregulated while 459 genes were down regulated by at least 2-fold when comparing the srpA mutant strain 1-9 to its parent strain PAK-AR2.These genes were classified into 21 major cellular processes based on the annotation of KEGG_B_class or further grouped into several major metabolic pathways, such as ribosomal proteins, type III secretion system (T3SS), type VI secretion system (T6SS), chemotaxis, cell motility, and cell shape control.More and more small proteins that were ignored from typical genome annotations have now been experimentally demonstrated to play important regulatory roles on various bacterial metabolic.
Project description:The alkaliphilic halotolerant bacterium Bacillus sp. N16-5 often faces salt stress in its natural habitats. One-color microarrays was used to investigate transcriptome expression profiles of Bacillus sp. N16-5 adaptation reactions to prolonged grown at different salinities (0%, 2%, 8% and 15% NaCl) and the initial reaction to suddenly alter salinity from 0% to 8% NaCl. Salt induced gene expression was measured when culture was grown on different salinities (0%, 2%, 8% and 15% NaCl) to mid-logarithmic phase. And salt induced gene expression was also measured at 0 min, 10 min, 30 min, 60min, 120min after a sudden change salinity from 0% to 8% NaCl.
Project description:RNA decay is central to gene regulation and ensures that once genes are expressed, they have a finite lifespan. In Escherichia coli, bulk degradation is carried out by the RNA degradosome, a multi-protein complex that is scaffolded by the endoribonuclease RNase E. RNase E recognises RNA substrates through at least four RNA-binding sites that feed RNA to the catalytic centre. The ribonuclease consists of a structured N-terminal domain that contains the catalytic site, and an intrinsically disordered C-terminal domain that contains two unstructured RNA-binding sites termed RBD and AR2. The intrinsically disordered domain provides a major pathway for RNA recruitment to the catalytic centre, including mRNAs targeted by regulatory small RNAs (sRNAs). To understand how sRNA-mRNA pairs are recognised by the intrinsically disordered C-terminal domain we have engineered protease cleavage sites that separate the AR2 microdomain from RNase E and performed UV-crosslinking and sequencing (termed split-CRAC). We find that the AR2 binding site recognises AUAA motifs within mRNA translation initiation regions (RBS and start codons) and is depleted of sRNA interactions. We demonstrate that the AUAA motif is required for sRNA regulation of the highly abundant murein lipoprotein, Lpp, and that mRNAs with an AR2 motif tend to be more stable and more highly expressed.
Project description:Alkaline hemicellulytic bacteria Bacillus sp. N16-5 has abroad substrate spectrum and exhibits great growth ability on complex carbohydrates. In order to get insight into its carbohydrate utilization mechanism, global transcriptional profiles were separately determined for growth on glucose, fructose, mannose, galactose, arabinose, xylose, galactomannan, xylan, pectin and carboxymethyl cellulose by using one-color microarrays.
