Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

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Query: EC:2.7.7.49 (reverse transcriptase)
31,746 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Legumes interact with nodulating bacteria that convert atmospheric nitrogen into ammonia for plant use. This nitrogen fixation takes place within root nodules that form after infection of root hairs by compatible rhizobia. Using cDNA microarrays, we monitored gene expression in soybean (Glycine max) inoculated with the nodulating bacterium Bradyrhizobium japonicum 4, 8, and 16 days after inoculation, timepoints that coincide with nodule development and the onset of nitrogen fixation. This experiment identified several thousand genes that were differentially expressed in response to B. japonicum inoculation. Expression of 27 genes was analyzed by quantitative reverse transcriptase-polymerase chain reaction, and their expression patterns mimicked the microarray results, confirming integrity of analyses. The microarray results suggest that B. japonicum reduces plant defense responses during nodule development. In addition, the data revealed a high level of regulatory complexity (transcriptional, post-transcriptional, translational, post-translational) that is likely essential for development of the symbiosis and adjustment to an altered nutritional status.
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PMID:Transcription profiling of soybean nodulation by Bradyrhizobium japonicum. 1839 23

An understanding of differential gene expression in highly metastatic osteosarcoma could provide gene targets for treatment of metastases. We compared gene expression profiles of high- (LM7) and low- (LM2) metastatic SaOS2-derived cell lines in an in vitro tissue culture model and examined several differentially regulated genes in vivo in a murine orthotopic xenograft model. We hypothesized an orthotopic inoculation of LM2 and LM7 cells would establish a primary lesion and the gene expression profile of cells grafted in this fashion would resemble the gene expression profile observed in an in vitro model. Thirty-five days after inoculation, animals were euthanized and both tibiae were harvested and rapidly frozen in liquid nitrogen. Human-specific GAPDH mRNA was present in two of four tibias inoculated with LM2 cells and three of four tibias inoculated with LM7 cells. Tibiae displaying the presence of human cells were assayed by semiquantitative reverse transcriptase polymerase chain reaction. We observed poor correspondence of in vitro to in vivo gene expression for either cell line. Accordingly, in vitro osteosarcoma gene expression data must be interpreted with caution until confirmed in vivo. Our orthotopic injection model allowed in vivo study of differential gene expression between these two cell lines but did not show radiographic evidence of an established primary lesion.
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PMID:Metastatic osteosarcoma gene expression differs in vitro and in vivo. 1851 56

The Anabaena genome contains two ORFs that appear to encode glutaminases. The genes were expressed as histidine-tagged fusion proteins in Escherichia coli. The purified proteins possessed glutaminase activity using l-glutamine as the substrate, but differed in biochemical properties. All2934 showed an optimal activity at 20 degrees C and pH 6.0, with a higher affinity for l-glutamine than All4774, which had optimal activity at 37 degrees C and pH 7.5. Remarkably, the glutaminase activity of All2934 was phosphate dependent, while All4774 was phosphate independent. The expression of all2934 and all4774 was analyzed using semi-quantitative reverse transcriptase-PCR. The expression level of all2934 was much higher than that of all4774 under normal and nitrogen-depletion conditions, indicating that All2934 may play an important role in metabolizing glutamine in Anabaena.
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PMID:Characterization of two glutaminases from the filamentous cyanobacterium Anabaena sp. PCC 7120. 1905 11

A simple, specific, and high throughput liquid chromatography tandem mass spectrometry method is developed for the determination of tenofovir, a nucleotide reverse transcriptase inhibitor, in human plasma using adefovir as internal standard. Plasma samples are prepared by solid-phase extraction of the analyte and internal standard using Waters Oasis MCX cartridges (1 cc, 30 mg). The chromatographic separation is achieved on a reversed-phase Chromolith, C18 analytical column (100 mmx4.6 mm, 5 microm) under isocratic conditions. The mobile phase consists of 0.5% formic acid in water and acetonitrile (90:10, v/v) to give a run time of 1.8 min. The protonated precursor-->product ion transitions for tenofovir and IS are monitored on a triple quadrupole mass spectrometer, operating in the multiple reaction monitoring and positive ion mode. The fragmentation pathways for tenofovir are studied by varying the collision energy (5-55 V) using nitrogen as CAD gas. A linear dynamic range of 3.1-1002.0 ng/mL is established using 0.2 mL plasma sample. The method is fully validated for its sensitivity, selectivity, accuracy and precision, matrix effect, recovery, stability, and dilution integrity. It is applied to a bioequivalence study in 43 human subjects after oral administration of 300 mg tablet formulation under fasting conditions.
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PMID:Rapid and specific liquid chromatographic tandem mass spectrometric determination of tenofovir in human plasma and its fragmentation study. 1922 22

The diazotrophic cyanobacterium Crocosphaera watsonii supplies fixed nitrogen (N) to N-depleted surface waters of the tropical oceans, but the factors that determine its distribution and contribution to global N(2) fixation are not well constrained for natural populations. Despite the heterogeneity of the marine environment, the genome of C. watsonii is highly conserved in nucleotide sequence in contrast to sympatric planktonic cyanobacteria. We applied a whole assemblage shotgun transcript sequencing approach to samples collected from a bloom of C. watsonii observed in the South Pacific to understand the genomic mechanisms that may lead to high population densities. We obtained 999 C. watsonii transcript reads from two metatranscriptomes prepared from mixed assemblage RNA collected in the day and at night. The C. watsonii population had unexpectedly high transcription of hypothetical protein genes (31% of protein-encoding genes) and transposases (12%). Furthermore, genes were expressed that are necessary for living in the oligotrophic ocean, including the nitrogenase cluster and the iron-stress-induced protein A (isiA) that functions to protect photosystem I from high-light-induced damage. C. watsonii transcripts retrieved from metatranscriptomes at other locations in the southwest Pacific Ocean, station ALOHA and the equatorial Atlantic Ocean were similar in composition to those recovered in the enriched population. Quantitative PCR and quantitative reverse transcriptase PCR were used to confirm the high expression of these genes within the bloom, but transcription patterns varied at shallower and deeper horizons. These data represent the first transcript study of a rare individual microorganism in situ and provide insight into the mechanisms of genome diversification and the ecophysiology of natural populations of keystone organisms that are important in global nitrogen cycling.
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PMID:In situ transcriptomic analysis of the globally important keystone N2-fixing taxon Crocosphaera watsonii. 1922 52

Oxidative stress is involved in memory impairment at high altitude (HA). The aim of the present study was to investigate the involvement of reactive nitrogen species in hippocampus, cortex and striatum of rat brain under simulated HA conditions. Rats were exposed to hypobaric hypoxia (HH) equivalent to 6100 m of HA in an animal decompression chamber for 3, 7, 14 and 21 days. Biochemical estimation of free radicals, nitric oxide (NO) level along with immunoreactivity, reverse transcriptase polymerase chain reaction (RT-PCR) and western blot of neuronal nitric oxide synthase (nNOS), neurodegeneration and DNA fragmentation were studied after HH exposure. The free radicals, NO level, nNOS immunoreactivity (nNOS-IR), nNOS expression, neurodegeneration and DNA fragmentation were increased in hippocampus, cortex and striatum after HH exposure. After 7 and 14 days of HH exposure, the nNOS-IR, nNOS expression, free radical, NO level, neurodegeneration and DNA fragmentation were increased in comparison to 3 or 21 days of HH. The NO system may be involved in increasing oxidative stress and neurodegeneration after HH.
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PMID:Nitric oxide system is involved in hypobaric hypoxia-induced oxidative stress in rat brain. 1942 54

Carbonic anhydrase (CA; [EC 4.2.1.1]) is a ubiquitous enzyme catalysing the reversible hydration of CO(2) to bicarbonate, a reaction that supports various biochemical and physiological functions. Genome analysis of Azospirillum brasilense, a nonphotosynthetic, nitrogen-fixing, rhizobacterium, revealed an ORF with homology to beta-class carbonic anhydrases (CAs). Biochemical characteristics of the beta-class CA of A. brasilense, analysed after cloning the gene (designated as bca), overexpressing in Escherichia coli and purifying the protein by affinity purification, revealed that the native recombinant enzyme is a homotetramer, inhibited by the known CA inhibitors. CA activity in A. brasilense cell extracts, reverse transcriptase (RT)-PCR and Western blot analyses showed that bca was constitutively expressed under aerobic conditions. Lower beta-galactosidase activity in A. brasilense cells harbouring bca promoter: lacZ fusion during the stationary phase or during growth on 3% CO(2) enriched air or at acidic pH indicated that the transcription of bca was downregulated by the stationary phase, elevated CO(2) levels and acidic pH conditions. These observations were also supported by RT-PCR analysis. Thus, beta-CA in A. brasilense seems to be required for scavenging CO(2) from the ambient air and the requirement of CO(2) hydration seems to be higher for the cultures growing exponentially at neutral to alkaline pH.
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PMID:Regulation of expression and biochemical characterization of a beta-class carbonic anhydrase from the plant growth-promoting rhizobacterium, Azospirillum brasilense Sp7. 1969 14

This study examined the question of whether deficiency of xCT, a cystine-transporter gene, exacerbates ischemia-reperfusion-induced acute renal failure (ARF). Two weeks after the right nephrectomy of male mice at 16-18weeks of age, the left renal vessels were clamped for 45min to induce renal ischemia. After (24h) induction of ischemia, xCT(-/-) mice had elevated concentrations of blood urea nitrogen and creatinine indicative of ARF, while in xCT(+/-) and xCT(+/+) mice, these parameters did not differ from the sham-operated mice. Immunohistochemical analyses of kidneys using antibodies against the oxidative stress markers revealed stronger staining in xCT(-/-) mice compared with xCT(+/+) mice. Induction of xCT mRNA in the kidneys of xCT(+/+) mice was demonstrated using reverse transcriptase (RT)-PCR analysis and was further confirmed using quantitative RT-PCR. These data provide the first in vivo evidence that xCT is induced by oxidative stress and helps prevent ischemia-reperfusion injury to kidneys.
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PMID:Aggravation of ischemia-reperfusion-triggered acute renal failure in xCT-deficient mice. 1969 16

Abstract A diverse assemblage of diazotrophic bacteria exists in the rhizosphere of the smooth cordgrass, Spartina alterniflora, but the taxa actively involved in nitrogen fixation have not been determined. In order to identify the diazotrophs that were actively expressing nifH, the gene encoding the nitrogenase iron protein, mRNA was extracted from Spartina rhizosphere samples and nifH-specific seminested reverse transcriptase-PCR performed. Expressed nifH sequences were recovered from organisms affiliated with the (gamma-+beta-) Proteobacteria and the anaerobes. Most of the expressed nifH sequences were highly similar (>/=95% similarity) to sequences previously recovered from Spartina rhizosphere DNA using conventional nifH-specific PCR. These sequences were also similar, although not identical to the nifH sequences of Pseudomonas stutzeri, Vibrio diazotrophicus, Desulfovibrio africanus, and Desulfovibrio gigas.
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PMID:Expression of nifH genes by diazotrophic bacteria in the rhizosphere of short form Spartina alterniflora. 1971 72

Phosphorus (P) deficiency is widespread in regions where the common bean (Phaseolus vulgaris), the most important legume for human consumption, is produced, and it is perhaps the factor that most limits nitrogen fixation. Global gene expression and metabolome approaches were used to investigate the responses of nodules from common bean plants inoculated with Rhizobium tropici CIAT899 grown under P-deficient and P-sufficient conditions. P-deficient inoculated plants showed drastic reduction in nodulation and nitrogenase activity as determined by acetylene reduction assay. Nodule transcript profiling was performed through hybridization of nylon filter arrays spotted with cDNAs, approximately 4,000 unigene set, from the nodule and P-deficient root library. A total of 459 genes, representing different biological processes according to updated annotation using the UniProt Knowledgebase database, showed significant differential expression in response to P: 59% of these were induced in P-deficient nodules. The expression platform for transcription factor genes based in quantitative reverse transcriptase-polymerase chain reaction revealed that 37 transcription factor genes were differentially expressed in P-deficient nodules and only one gene was repressed. Data from nontargeted metabolic profiles indicated that amino acids and other nitrogen metabolites were decreased, while organic and polyhydroxy acids were accumulated, in P-deficient nodules. Bioinformatics analyses using MapMan and PathExpress software tools, customized to common bean, were utilized for the analysis of global changes in gene expression that affected overall metabolism. Glycolysis and glycerolipid metabolism, and starch and Suc metabolism, were identified among the pathways significantly induced or repressed in P-deficient nodules, respectively.
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PMID:Global changes in the transcript and metabolic profiles during symbiotic nitrogen fixation in phosphorus-stressed common bean plants. 1975 43


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