Catalytic abilities of the Schistosoma mansoni hammerhead ribozyme with mutated substrates in ice

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Calvird, Audrey
Wartell, Roger M.
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The synthesis of biomolecules in an environment similar to a pre-biotic Earth within the field of evolutionary chemistry has applications in understanding abiogenesis and the evolution of early biological systems on Earth. The RNA world is a prebiotic environment proposed and subsequently heavily studied in effort to better understand how biochemical reactions started on Earth. This study proposes to assess the catalytic ability of a Hammerhead Ribozyme (HHRz) isolated from the protozoa Schistosoma mansoni (Schist HHRz). It has been previously shown that the Schist HHRz can catalyze the synthesis of extended sequences of ribonucleic acid (RNA) by ligating two RNA substrates in ice, as well as cleave the same substrate back to the original separate substrates in the presence of Mg2+ (Lie et al. 2016). The same study showed that the Schist HHRz is able to catalyze the ligation with mutated substrates (Lie et al. 2016). This study aims to evaluate Schist HHRz’s ability to ligate a mutated substrate in comparison to the wild type substrate in ice, as well as compare the kinetics of the Schist HHRz to cleave the mutated and wild type substrates in the presence of Mg2+. This study will utilize the techniques specified by Lie et al. (2016) to further investigate the extent of the ligation of a mutated substrate catalyzed by the Schist HHRz compared to that of the wild type substrate. This study also assays the kinetics of the cleavage reaction catalyzed by the same Schist HHRz of both the wild type substrate and mutated substrate. This study hopes to improve the understanding of pre-biotic biochemistry in efforts to identify possible mechanisms or models for RNA polymerization by ribozymes on early Earth.
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