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Mechanism-Driven Approaches And Novel Constructs For High Purity Rna Synthesis, Kithmie Malagodapathiranage
Mechanism-Driven Approaches And Novel Constructs For High Purity Rna Synthesis, Kithmie Malagodapathiranage
Doctoral Dissertations
RNA is poised to revolutionize medicine. By simply changing the sequence, one therapeutic can be converted into a wholly new one, with little or no change in manufacturing and formulation. While a single mRNA vaccine produced at massive scale can treat billions, the re-codability of RNA will also enable the widespread growth of personalized medicines. T7 RNA polymerase is highly efficient at the synthesis of therapeutic RNA, but is known to produce unintended RNA impurities during synthesis. These products arise from the encoded RNA rebinding the enzyme such that its 3’ end serves as a primer for extension. This leads …
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 …
In Vitro Enzymatic Assay Of Rna Methylation, Pamela Jean Eubanks Gallup
In Vitro Enzymatic Assay Of Rna Methylation, Pamela Jean Eubanks Gallup
Chemistry & Biochemistry Theses & Dissertations
An assay procedure for in vitro enzymatic methylation of mannnalian ribosomal RNA has been developed in this study. The assay procedure, utilized for the comparison of normal and neoplastic methylase activities (using mouse liver and Ehrlich ascites cells as sources of enzyme), is a modification of previously published methods (52,53). A 100,000 x g supernatant (SlOO) enzyme preparation was incubated with 28S-5.8S rRNA and tritium-labeled S-adenosyl-L-methionine. The RNA was extracted, applied to DEAE cellulose paper, washed, and the radioactivity counted. The neoplastic cell methylase preparation was more active in methylating both exogenous neoplastic and normal 28S-5.8S rRNA than the normal …