Project description:The study aims to identify the metabolic differences between two promising fast-growing, non-model cyanobacterial strains, S. elongatus PCC 11801 and PCC 11802. To this end, dynamic 13C-labeling experiments were carried out in the two cyanobacterial strains grown in shake flasks at a similar light intensity of approx. 300-350 µmole photons.m-2. s-1. The samples for metabolomics analysis were collected during the exponential growth phase at an optical cell density of 0.5-0.6. The detailed protocol for experiment can be found in the protocol file.
Project description:The study aims to identify the metabolic differences between two promising fast-growing, non-model cyanobacterial strains, S. elongatus PCC 11801 and PCC 11802. To this end, experiments were carried out to measure metabolite levels in the two cyanobacterial strains grown in shake flasks at a similar light intensity of approx. 300-350 µmole photons.m-2. s-1. The samples for metabolomics analysis were collected during the exponential growth phase at an optical cell density of 0.5-0.6. Isotopic ratio method was utilized to compare the metabolite levels and delineate the differences in their metabolic pathways.
Project description:We installed and optimized a genetic tool that allows control over protein stability in a model cyanobacterium. This tool has potential uses for the fundamental study of cyanobacterial genes, and may be useful for the design of more sophisticated, bioindustrial cyanobacterial strains.
Project description:Investigation of whole genome gene expression level changes in two strains of the cyanobacteria Atelocyanobacterium thalasaa (UCYN-A) from environmental samples. The diel gene expression analyzed in this study is further described in Muñoz-Marin, M., I. N. Shilova, T. Shi, H. Farnelid & J. P. Zehr. 2017. Unicellular cyanobacterial symbiosis facilitates aerobic nitrogen fixation. Science (to be submitted).
Project description:Changes in cellular metabolism contribute to the development and progression of tumors, and can render tumors vulnerable to interventions. However, studies of human cancer metabolism remain limited due to technical challenges of detecting and quantifying small molecules, the highly interconnected nature of metabolic pathways, and the lack of designated tools to analyze and integrate metabolomics with other âomics data. Our study generates the largest comprehensive metabolomics dataset on a single cancer type, and provides a significant advance in integration of metabolomics with sequencing data. Our results highlight the massive re-organization of cellular metabolism as tumors progress and acquire more aggressive features. The results of our work are made available through an interactive public data portal for cancer research community. 10 RNA samples from human ccRCC tumors analyzed from the high glutathione cluster
Project description:20 different cyanobacterial strains were cultivated in shaking flasks and harvested by centrifugation. The supernatants were extracted via solid phase extraction (SPE), the cell pellets with 80% MeOH.
Project description:Cyanobacteria are phototrophic prokaryotes that can convert inorganic carbon as CO2 into organic carbon compounds at the expense of light energy. In addition, they need only a few inorganic nutrients and can be cultivated in high densities using non-arable land and seawater. This features qualified cyanobacteria as attractive organisms for the production of third generation biofuels as part of the development of future CO2-neutral energy production. Synechocystis sp. PCC 6803 represents one of the most widely used cyanobacterial model strains. On the basis of its available genome sequence and genetic tools, many strains of Synechocystis have been generated that produce different biotechnological products. Efficient isoprene production is an attractive goal, since this compound represents not only an energy-rich biofuel but is also used as chemical feedstock. Here, we report on our attempts to generate isoprene-producing strains of Synechocystis. The cDNA of a codon-optimized plant isoprene synthase (IspS) was cloned under the control of different Synechocystis promoters, which ensure strong constitutive or light-regulated ispS expression. The expression of the ispS gene was quantified by qPCR, whereas the amount of isoprene was quantified using GC-MS. Incubation of our strains at different salt conditions had marked impact on the isoprene production rates. Under low salt conditions, a good correlation was found between ispS expression and isoprene production rate. However, the cultivation of isoprene production strains under salt-supplemented conditions decreased isoprene production despite the fact that ispS expression was salt-stimulated. The characterization of the metabolome of isoprene producing strains indicated that isoprene production might be limited by insufficient precursor levels. Our isoprene production rates under low salt conditions were 2 - 6.5times higher compared to the previous report of Lindberg et al. (2010). These results can be used to guide future attempts establishing the isoprene production with cyanobacterial host systems.