Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.1.3.1 (alkaline phosphatase)
 47,916 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

This protocol describes the purification by denaturing polyacrylamide gel electrophoresis of RNA linkers for cross-linking immunoprecipitation (CLIP). Purification is necessary because if the 3' linker loses the puromycin blocking group, concatemerization of the 3' linker will occur during the 3' linker ligation reaction. In addition, truncated linkers make bioinformatic processing of the sequencing results more difficult than it need be. Additionally, this protocol describes the treatment of coimmunoprecipitated RNA tags for CLIP with alkaline phosphatase to remove the 3' phosphate remaining after RNase digestion. Dephosphorylation prevents intramolecular circularization of RNA during subsequent ligation to the linker. The purified RNA linker, blocked with puromycin at its 3' end to prevent linker-linker multimerization, is then ligated to the 3' end of the RNA tag. Removal of free linker is accomplished by performing the ligation while the RNABP:RNA complex is associated, via antibody, to protein A Dynabeads, allowing thorough washing and linker removal. Additional purification is achieved by SDS-PAGE and transfer of the size-selected RNABP:RNA complexes to nitrocellulose.
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PMID:3'-Linker Ligation and Size Selection by SDS-PAGE for Cross-Linking Immunoprecipitation (CLIP). 3051 Jan 27

The aim of experiments was to investigate a maximal efficiency of PSII, as a marker indicating growth, vigor, energetic value and physiological activity of sorghum fertilized with wastes from a biomass biodigestion to methane in a distillery integrated with a biogas plant using corn grains as substrate. The sorghum plants grown outdoor in different climate and in pots and in field were fertilized with different doses of the waste or Apol-humus - a soil improver and Stymjod - a nano-organic-mineral fertilizer. The maximal efficiency of PSII, in comparison with plant growth and health, chlorophyll content, gas exchange, activity of selected enzymes, element content in leaves and energetic value were studied. The wastes applied to soil resulted in increased maximal efficiency of PSII and the doses of 30 m3 ha-1 and 40-50 m3 ha-1 of the non-centrifuged and centrifuged ones, respectively, were most efficient. This enhancement was associated with the increased kinetics of plant growth, their health, fresh and dry biomass and physiological activity of plants as evidenced by activity of acid and alkaline phosphatase, RNase and dehydrogenase, as well as by gas exchange: net photosynthesis, transpiration, stomatal conductance, intercellular CO2 concentration and index of chlorophyll content in leaves. The fertilization with Apol-humus and Stymjod additionally increased maximal photochemical efficiency of PSII and plant development, biomass yield and physiological activity. The results indicate that waste from a biomass biodigestion to methane can be used as a natural fertilizer in sorghum crops and this ensures their recycling and environmental protection. The measurement values of maximal efficiency of PSII were proportionally to the vigor, growth and physiological activity of the plants. The obtained results indicate that the maximal efficiency of PSII in sorghum plants is a non-destructive method for defining the degree of growth and may be used as a marker of plant vigor and health, development and physiological activity expressed by gas exchange and activity of selected enzymes.
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PMID:Maximal Efficiency of PSII as a Marker of Sorghum Development Fertilized With Waste From a Biomass Biodigestion to Methane. 3067 Oct 72


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