Interactions of SARS-CoV-2 N-protein

点击翻译按钮获取中文摘要

To allow an immediate start of our investigations we have examined N-protein from several commercial sources and identified suitable protein constructs. Simultaneously we have started to develop in-house protein expression, jointly with the laboratory of Dr. Eugene Valkov. We are currently optimizing solution conditions, and are exploring and implementing purification strategies. We have carried out initial sedimentation velocity analytical ultracentrifugation experiments to assess the oligomeric state and found it to be dimeric, similar to SARS-CoV N-protein. We explored different fluorescent labeling strategies to better measure the affinity of self-association. We have also carried out initial binding experiments with small oligonucleotides, which will help us to optimize the biophysical assays for assembly reactions. In collaboration with Dr. Greg Piszczek we have begun to characterize the thermal properties of the protein and its complexes. We are evaluating the first results.