Project description:MKN-45 cells were treated with PBS, BF-TK, BF/GCV or BF-TK/GCV for 48 h (GCV, 167 µg/ml), respectively. BF-TK (n = 3), BF/GCV (n = 3) or BF-TK/GCV (n = 3) groups were analyzed in triplicate.
Project description:Foldon (F) is a specific partial sequence from the C-terminal domain of T4 fibritin, which is found at the neck of the T4 bacteriophage. The amino acid sequence (27 residues) in single letter code is GYIPEAPRDGQAYVRKDGEWVLLSTFL. In neutral solution (pH 6.9) it forms a homo-trimer (F-F-F). Biotinylated foldon (bF) is composed of the exact same amino acid sequence but contains a biotin residue attached to the N-terminal amino acid residue via two PEG linker molecules. In solution with neutral pH it forms a homo-trimer (bF-bF-bF). Mixing of both homo-trimers in solution with neutral pH spontaneously produces hetero-trimers (F-F-bF and F-bF-bF).
Project description:The adult mammalian brain is composed of distinct regions that have specialized roles. The BF/POA regions are thought to have an important role in the regulation of sleep/wake behavior. However, genetic markers of the responsible cells for the regulation of sleep/wake behavior are largely unknown. To identify the molecular markers of the BF/POA regions, we sampled the BF/POA regions and compared gene expression in the BF/POA regions with those of other brain regions which we previously reported in the BrainStars (B*) project, in which we sampled ~50 small brain regions, including sensory centers and centers for motion, time, memory, fear, and feeding. We sampled each region every 4 hours for 24 hours, and pooled the sample sets for DNA-microarray assays. We then used informatics to identify candidates for genes with high or low expression in the BF/POA regions. We used our findings to develop an integrated database (http://poabf.brainstars.org/) for exploring genome-wide expression in the adult mouse brain including the BF/POA regions.
Project description:We have shown that pre-incubation with Bifidobacterium bifidum (B. bifidum) strain BF-1, a probiotic strain known to improve H. pylori-associated gastritis, suppresses the induction of IL-8 by the pathogen. To investigate how BF-1 affects gene expression in H. pylori-infected cells, we performed microarray analysis to assess gene expression in epithelial cells, which had been pre-incubated with BF-1 and infected with H. pylori.
Project description:Gut-educated IgA-secreting plasma cells that disseminate beyond the mucosa and into systemic tissues can help prevent disease in several contexts. Here we show, the commensal bacteria Bacteroides fragilis (Bf), is an efficient inducer of systemic IgA responses. The generation of bone marrow IgA plasma cells and high levels of serum IgA specific to Bf requires robust intestinal colonization. Bf-specific IgA responses were severely diminished in mice lacking Peyer’s patches, but not mice lacking a cecal patch. Colonization resulted in few changes in the host transcriptional profile in the gut, suggesting a commensal relationship. High levels of Bf-specific serum IgA, but not IgG, provided protection from peritoneal abscess formation in a bowel perforation model of Bf dissemination. These findings demonstrate a critical role for bacterial colonization and Peyer’s patches in the induction of robust systemic IgA responses that confer protection from bacterial dissemination originating from the gut.
Project description:Our goal is to convert methane efficiently into liquid fuels that may be more readily transported. Since aerobic oxidation of methane is less efficient, we focused on anaerobic processes to capture methane, which are accomplished by anaerobic methanotrophic archaea (ANME) in consortia. However, no pure culture capable of oxidizing and growing on methane anaerobically has been isolated. In this study, Methanosarcina acetivorans, an archaeal methanogen, was metabolically engineered to take up methane, rather than to generate it. To capture methane, we cloned the DNA coding for the enzyme methyl-coenzyme M reductase (Mcr) from an unculturable archaeal organism from a Black Sea mat into M. acetivorans to effectively run methanogenesis in reverse. The engineered strain produces primarily acetate, and our results demonstrate that pure cultures can grow anaerobically on methane.
Project description:RNA-Seq profiling of Methylomicrobium alcaliphilum strain 20Z grown in batch on methane. The RNA-Seq work is one part of a systems approach to characterizing metabolism of 20Z during growth on methane. We demonstrate that methane assimilation is coupled with a highly efficient pyrophosphate-mediated glycolytic pathway, which under O2 limitation participates in a novel form of fermentation-based methanotrophy. This surprising discovery suggests a novel mode of methane utilization in oxygen-limited environments, and opens new opportunities for a modular approach towards producing a variety of excreted chemical products using methane as a feedstock.
Project description:The BF-3 pocket of the androgen receptor (AR) has been identified as an allosteric modulator of the transactivation function of the AR. We now demonstrate that a duplicated GARRPR motif at the N-terminus of the cochaperone Bag-1L functions through this BF-3 domain. Amino acid exchanges in these two motifs impair binding of Bag-1L to the AR but increase the androgen-dependent activation of a subset of AR-target genes. We have therefore identified GARRPR as a novel BF-3 regulatory sequence important for fine-tuning the activity of the receptor.
Project description:RNA-Seq profiling of Methylomicrobium alcaliphilum strain 20Z grown in batch on methane. The RNA-Seq work is one part of a systems approach to characterizing metabolism of 20Z during growth on methane. We demonstrate that methane assimilation is coupled with a highly efficient pyrophosphate-mediated glycolytic pathway, which under O2 limitation participates in a novel form of fermentation-based methanotrophy. This surprising discovery suggests a novel mode of methane utilization in oxygen-limited environments, and opens new opportunities for a modular approach towards producing a variety of excreted chemical products using methane as a feedstock. Four replicates of batch growth
Project description:Comparative transcriptomic analysis between methane- and methane plus xylose- grown cultures revealed different transcriptional responses of pXyl M. alcaliphilum 20Z strain on diffirent growth modes.