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UBC Theses and Dissertations

Differences in the regulation of translation dynamics in eukaryotic cell-free protein synthesis systems Khouhak, Jalal


Cell-free protein synthesis systems (CFPSS) constitute a powerful tool to study translation dynamics. Yet, they can produce various complex dynamics across conditions and systems that can be challenging to explain. To investigate and interpret differences between eukaryotic CFPSS, we studied two lysate systems; one from Rabbit Reticulocyte and the other from Plasmodium Falciparum. Additionally, we measured the level of protein production from a sequence expressing nanoluciferase over time under various conditions. Upon the addition of mRNA at varied concentrations and pre-incubation times, we observed that the two systems exhibit distinct dynamics, which are quantified and interpreted as the result of dissimilar reactions. In the Rabbit system, translation reaction establishes quickly, but the system is sensitive to a translation-independent degradation of resources. Conversely, the P. falciparum system exhibits a significant delay in initiating translation, while no significant impact of pre-incubation time was detected. Furthermore, we introduced a system of differential equations to mathematically model the translation dynamics, as a function of the mRNA concentration and available resources. Upon fitting the model parameters to the experimental data, the model effectively captured the dynamics of both systems. We observed a notable difference in the transition rate from inactive to active mRNA between Rabbit and P. falciparum, which highlights the different key reactions and limiting factors that regulate translation across eukaryotic CFPSS.

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