Project description:In Nannochloropsis oceanica IMET1, transcript knockdown of a cytosolic carbonic anhydrase (CA2; g2018) specifically inhibited by HC resulted in ~45%, ~30% and ~40% elevation of photosynthetic oxygen evolution rate, growth rate and biomass accumulation rate under high CO2 (5% ), respectively. This CA2-knockdown mutant is demonated as M2. To probe mechanistic links underlying the mutant (M2; RNAi-knockdown line of carbonic anhydrase (CA2)) phenotypes, temporal transcriptomic profiles are compared between RNAi-knockdown line of carbonic anhydrase (CA2) and WT, at 12 h and 24h under high CO2 (5%).
Project description:geLC-MS/MS analysis was performed to identify proteins that are present in apoplastic fluid isolated from rosette leaves of 8-week-old wild type and carbonic anhydrase (ca1ca4) mutant Arabidopsis. Plants were grown in either low (150 ppm) or high (500 ppm) CO2.
Project description:Wild-type Brucella ovis ATCC 25840 requires the addition of 5% CO2 to the atmosphere to grow on either nutrient agar plates or in liquid broth culture. The goal of this study was to measure the transcriptional response of two Brucella ovis strains under high (5%) and low (0.04%) CO2. The two strains assayed were 1) wild-type and 2) a spontaneous mutant that can be cultivated in standard atmospheric levels of CO2 (~0.04%). Wild-type B. ovis harbors a single nucleotide insertion at the 3' end of a beta carbonic anhydrase gene, bcaA, which renders it non-functional. The spontaneous mutant lost this nucleotide insertion, which restored the consensus reading frame and results in a functional BcaA protein.
Project description:cea05-01_carbonic-anhydrase - carbonic anhydrase (1 or 2) simple or double mutants - Identification of genes differentially expressed in leaves of single CA1 and CA2 T-DNA insertional mutants and in the corresponding double mutant vs wild type - CA1 (At3g01500) and CA2 (At5g14740) T-DNA insertional mutant lines, the double (CA1+CA2) mutant and wild type Arabidopsis seeds were sown in soil in a phytotron. Leaves were harvested 40 days later for RNA extraction
Project description:carbonic anhydrase (1 or 2) simple or double mutants-Comparative transcriptome analysis between carbonic anhydrase mutants and wild type Arabidopsis
Project description:Physiological and molecular evidences have shown earlier that low CO2 might have been a major driving force during the evolution of C4 photosynthesis, not only being a selection pressure but also as a signaling agent. However, a mechanistic linkage between low CO2 and C4 emergence is missing. In this study, using transcriptomics study in model plant Arabidopsis thaliana, we demonstrated that under long-term low CO2 treatments, the up-regulation of C4 related genes were linked to the up-regulation of genes involved in photorespiration, nitrogen assimilation and glycolysis. Plants under low CO2 also showed altered d13C. The carbonic anhydrase (CA), phosphoenolpyruvate carboxylase (PEPC), malate dehydrogenase (MDH), glutamine oxoglutarate aminotransferase (GS-GOGAT), which are required to reassimilate ammonia, were up-regulated under low CO2. Furthermore, under low CO2, genes involved in PEP regeneration from glycolysis were up-regulated while pyruvate orthophosphate dikinase (PPDK) was down-regulated, suggesting a route of PEP regeneration from glycolysis. All these results suggested that under long-term low CO2 treatment, the selection pressure to recapture the released ammonia from the increased photorespiration might have promoted the evolution of mechanisms for generation PEP from glycolysis and enhancement of enzymes catalyzing formation of oxaloacetate, one intermediate which can accept ammonia residue. These adjustments in metabolism provide a mechanistic linkage between low CO2 and evolution of C4 photosynthesis.
Project description:cea05-01_carbonic-anhydrase - carbonic anhydrase (1 or 2) simple or double mutants - Identification of genes differentially expressed in leaves of single CA1 and CA2 T-DNA insertional mutants and in the corresponding double mutant vs wild type - CA1 (At3g01500) and CA2 (At5g14740) T-DNA insertional mutant lines, the double (CA1+CA2) mutant and wild type Arabidopsis seeds were sown in soil in a phytotron. Leaves were harvested 40 days later for RNA extraction 6 dye-swap - gene knock out
Project description:Carbonic anhydrase IX (CA 9) is a transmembrane isoform of carbonic anhydrase (CA) that contributes to an acidification of tumor microenvironment. The expression of CA 9 in cervical tumors was shown to be strongly involved in high incidence of metastasis and poor prognosis. To search for the key regulators of metastasis related to ectopic expression of CA 9, we investigated differentially expressed gene profiles in CA 9- transfected cervix carcinoma cell line C33-A (CF) compared with mock-transfected (CP) cell line, using Affymetrix Human Genome U133 Plus 2.0 Array.
Project description:Carbonic anhydrase IX (CA 9) is a transmembrane isoform of carbonic anhydrase (CA) that contributes to an acidification of tumor microenvironment. The expression of CA 9 in cervical tumors was shown to be strongly involved in high incidence of metastasis and poor prognosis. To search for the key regulators of metastasis related to ectopic expression of CA 9, we investigated differentially expressed gene profiles in CA 9- transfected cervix carcinoma cell line C33-A (CF) compared with mock-transfected (CP) cell line, using Affymetrix Human Genome U133 Plus 2.0 Array. CF and CP stable cell-lines trasfected respectively with full-length human CA 9 cDNA cloned into the vector pcDNA3.0 and empty vector control were used for RNA extraction and hybridization on affymetrix microarrays.
Project description:Physiological effects of carbon dioxide and impact on genome-wide transcript profiles were analysed in chemostat cultures of Saccharomyces cerevisiae. In anaerobic, glucose-limited chemostat cultures grown at atmospheric pressure, cultivation under CO2-saturated conditions had only a marginal (<10%) impact on the biomass yield. Conversely, a 25% decrease of the biomass yield was found in aerobic, glucose-limited chemostat cultures aerated with a mixture of 79% CO2 and 21% O2. This observation indicated that respiratory metabolism is more sensitive to CO2 than fermentative metabolism. Consistent with the more pronounced physiological effects of CO2 in respiratory cultures, the number of CO2-responsive transcripts was higher in aerobic cultures than in anaerobic cultures. Many genes involved in mitochondrial functions showed a transcriptional response to elevated CO2 concentrations. This is consistent with an uncoupling effect of CO2 and/or intracellular bicarbonate on the mitochondrial inner membrane. Other transcripts that showed a significant transcriptional response to elevated CO2 included NCE103 (probably encoding carbonic anhydrase), PCK1 (encoding PEP carboxykinase) and members of the IMD gene family (encoding isozymes of inosine monophosphate dehydrogenase Keywords: Dose reponse
