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Full-Text Articles in Nucleic Acids, Nucleotides, and Nucleosides
A Purification And In Vitro Activity Assay For A (P)Ppgpp Synthetase From Clostridium Difficile, Astha Pokhrel, Asia Poudel, Erin B. Purcell
A Purification And In Vitro Activity Assay For A (P)Ppgpp Synthetase From Clostridium Difficile, Astha Pokhrel, Asia Poudel, Erin B. Purcell
Chemistry & Biochemistry Faculty Publications
Kinase and pyrophosphokinase enzymes transfer the gamma phosphate or the beta-gamma pyrophosphate moiety from nucleotide triphosphate precursors to substrates to create phosphorylated products. The use of γ-32-P labeled NTP precursors allows simultaneous monitoring of substrate utilization and product formation by radiography. Thin layer chromatography (TLC) on cellulose plates allows rapid separation and sensitive quantification of substrate and product. We present a method for utilizing the thin-layer chromatography to assay the pyrophosphokinase activity of a purified (p)ppGpp synthetase. This method has previously been used to characterize the activity of cyclic nucleotide and dinucleotide synthetases and is broadly suitable for …
Investigation Of Complex Formation By Oligomers Of Cytosine And Guanosine, Steven Roberts Davis
Investigation Of Complex Formation By Oligomers Of Cytosine And Guanosine, Steven Roberts Davis
Chemistry & Biochemistry Theses & Dissertations
Duplex formation between oligo(C:G) n where n=3 to 4 was shown not to occur under conditions favorable for duplex formation between poly G and poly C. Instead, a stable guano sine self-structure was found to form which a Tm of 50°C for (Gp)3 and 80°C for (Gp)4 at strand concentrations of 10-5M in 1M NaCl. Neither a duplex nor a self-structure formed in the absence of salt.
Oligomers of guanosine and cytosine were obtained by basic hydrolysis and separated according to chain length using DEAE Sephadex column chromatography. Separation of cytosine oligomers with chain lengths …