Application Of DNA Qrigami And Mechanism Research On Prion Assembly

Self-assembly,quite universal in nature,is an important phenomenon in super molecular chemistry.For bio-macromolecules like nucleic acids and proteins,people have explored a lot to give much information about the self-assembly mechanism of DNA/RNA,the primary structure of proteins,and protein interactions.Nevertheless,how to take advantage of DNA materials or what's the real mechanism of protein assembly haven't receive enough devotion.In this thesis,application of DNA origami and assembly mechanism of prion were studied.The specific research works are as follows:(1)With DNA origami,we developed a multiplexed DNA detection strategy based on positional encoding/decoding with self-assembled DNA nanostructures.The simple encoding/decoding method,PCR-free procedure and 2 uL-required sample volume make it to be an easy,fast and efficient DNA detection strategy.Besides,we also constructed DNA polygonal cavities with tunable shapes and sizes by DNA origami.Monomers with controlled slope on one end and flexible sticky ends could assemble in one-pot from ssDNAs to cavities,while most of ssDNAs were able to be reutilized.(2)We obtained the monomers of Sup35-NM and Sup35 and fibrils of Sup35-NM prion.By comparing the results of Sup35-NM and Sup35,we found the C terminal of Sup35 had negative effects on the formation of prions.Moreover,two kinds of proteins interacted with Sup35 were found for the first time during the purification procedure,which would make sense for heterogenous Sup35 studies.