Project description:A total gene expression approach was applied to study the methylotrophic nature of B. methanolicus by comparing the gene expression in bacteria grown methylotropic compared to non-methylotrophic. Genes of interest with different gene expression were quantified in the same RNA samples by real-time PCR, confirming the results found in the microarray experiment. Genes of special interest that are expressed higher when grown methylotrophic, were the RuMP pathway genes located on the pBM19.
Project description:A total gene expression approach was applied to study the methylotrophic nature of B. methanolicus by comparing the gene expression in bacteria grown methylotropic compared to non-methylotrophic. Genes of interest with different gene expression were quantified in the same RNA samples by real-time PCR, confirming the results found in the microarray experiment. Genes of special interest that are expressed higher when grown methylotrophic, were the RuMP pathway genes located on the pBM19. Bacillus methanolicus was grown in minimal media with either methanol or mannitol as carbon source. The experiment was preformed in triplicate, with bacterial cultures grown on 3 different days.
Project description:The mucus and cell-associated bacteria were isolated as a single fraction from ileal loops twelve hours post inoculation by scraping the epithelial surface of the intestine with a disposable plastic cell scraper after the loops had been cut open and gently rinsed in PBS buffer to wash away remaining luminal fluid. The mucus gel/cell associated fraction was suspended in Trizol and frozen on dry ice. Total RNA, which includes RNA from the infecting bacteria and from the host (see below), was isolated from the thawed Trizol suspension, treated with DNaseI (Applied Biosystems, Austin, TX) and cleaned by using the RNeasy kit (Qiagen, Valencia, CA).
Project description:The mucus and cell-associated bacteria were isolated as a single fraction from ileal loops eight hours post inoculation by scraping the epithelial surface of the intestine with a disposable plastic cell scraper after the loops had been cut open and gently rinsed in PBS buffer to wash away remaining luminal fluid. The mucus gel/cell associated fraction was suspended in Trizol and frozen on dry ice. Total RNA, which includes RNA from the infecting bacteria and from the host (see below), was isolated from the thawed Trizol suspension, treated with DNaseI (Applied Biosystems, Austin, TX) and cleaned by using the RNeasy kit (Qiagen, Valencia, CA).
Project description:<p>Four species of phytoplankton representing important bloom-forming species from three globally important phyla (Bacillariophyta, Haptophyta, and Ochrophyte) were cultured in this study. These species include the cosmopolitan diatom <em>Chaetoceros affinis</em> CCMP159 (isolated from Great South Bay, NY, USA, 1958), the haptophytes<em> Chrysochromulina polylepis </em>CCMP1757 (isolated from the North Sea 1988) and <em>Gephyrocapsa oceanica</em> RCC1303 (isolated from Arachon Bay, France, Jan 1999), and the raphidophyte <em>Heterosigma akashiwo </em>strain CCMP 2393 (isolated from Rehoboth Bay, Delaware, USA). Cultures were grown under three conditions: nitrogen-stress, phosphorus-stress, and replete conditions. Intracellular metabolites were extracted from cultures and analyzed with targeted and untargeted mass spectrometry-based metabolomics methods.</p>
Project description:The recent discovery of the lanthanide(Ln)-dependent methanol dehydrogenase (MDH) XoxF has expanded the spectrum of bacteria recognized for methylotrophic metabolism. Many bacteria, including rhizobia, have long escaped being categorized as methylotrophs because they exclusively produce XoxF-type Ln-dependent MDH and entirely lack the long-studied calcium-dependent MDH MxaFI. We report that the XoxF-type Ln-dependent MDH encoded by the smb20173 gene is the sole MDH that supports methylotrophic growth of Sinorhizobium meliloti. The Ln that consistently supported growth of S. meliloti in minimal media with methanol included lanthanum, cerium, praseodymium, and neodymium. Based on genome, whole-transcriptome, and mutant phenotype analyses, we propose a metabolic model for Ln-dependent methylotrophy in S. meliloti wherein oxidation of one-carbon compounds such as methanol generates the reducing power needed to assimilate carbon via the Calvin-Benson-Bassham cycle. By investigating how these newfound insights about Ln reshape our understanding of the methylotrophic capabilities of rhizobia, we explored how methanol produced by plants has the potential to create a nutritional niche in the rhizosphere. Using a Medicago sativa (alfalfa) nodule occupation assay, we found that the xoxF mutant strain was outcompeted by the wild-type strain only when Ln were available, suggesting that Ln-dependent methylotrophy is a potential nutritional mediator of the rhizobia-legume symbiosis.
Project description:The goal of this project was to compare the metabolite profiles of the: mouse gastric antrum and the mouse gastric corpus, the mouse gastric antrum and the mouse gastric antrum isolated glands, and the mouse gastric corpus and the mouse gastric corpus isolated glands.
2018-08-02 | MSV000082700 | GNPS
Project description:Genomes of novel bacteria isolated from Blanes Bay