Explore The Expression Purification And Interaction Of SARS-CoV-2 M Protein RIG-I And MAVS Protein

After the outbreak of the new coronavirus in 2019,it immediately attracted great attention from all over the world.The life sciences and medical circles have carried out all aspects of research on this new coronavirus.Studies have shown that the genome of the new coronavirus is a single-stranded RNA,while the RIG-I-like receptor in the innate immune system mainly recognizes RNA produced during virus replication in the cytoplasm.When the new coronavirus attacks the human body,The first to open the defense mechanism is the innate immune system.The innate immune system can not only detect pathogens at an early stage but also inhibit the growth of pathogens.This paper mainly studies the expression and purification of the SARS-CoV-2membrane(M)protein and the RIG-I-like receptor of the innate immune system RIG-I protein and MAVS protein,and explores whether these three proteins can interact with each other.In this paper,mammalian eukaryotic expression system combined with gel filtration chromatography is used to express and purify SARS-CoV-2 membrane(M)protein and MAVS protein,and E.coli prokaryotic expression system combined with gel filtration chromatography is used to purify RIG-I protein,and then use the pull down method and the direct mixing of the protein to pass through gel filtration chromatography to explore whether there is a direct interaction between these proteins.The results show that this paper has successfully mastered the construction method of the expression vector of SARS-CoV-2 membrane(M)protein and RIG-I protein,and has successfully purified the SARS-CoV-2 membrane(M)protein and RIG-I protein,but MAVS protein has not been purified yet.The SARS-CoV-2membrane(M)protein and RIG-I protein were pulled down,and The SARS-CoV-2membrane(M)protein and RIG-I protein were directly mixed and incubated,showed that the two proteins did not bind.In summary,this article carried out the expression and purification of SARS-CoV-2 membrane(M)protein and RIG-I protein,and initially explored whether SARS-CoV-2 membrane(M)protein and RIG-I protein can interact This can provide certain ideas for future research on the pathogenesis of the new coronavirus,and it can also provide help for the exploration of the structure of other types of coronaviruses.