Study On The Molecular Interaction Mechanism Between Cucumber Mosaic Virus 2b Protein And Compound LP1

Cucumber mosaic virus(CMV),which leads to cucumber mosaic virus disease,pepper mosaic virus disease and tobacco mosaic virus disease,causes huge economic losses to economic crops.At present,no effective anti-CMV agents to control the related viral disease Therefore,discovering CMV target protein is the key point to development anti-CMV agents.Hence,the discovery of potential molecular targets of CMV is of great significance for the discovery and development of novel anti-CMV agents.In this paper,CMV 2b protein was used as the potential molecular target and obtained by prokaryotic expression.On the one hand,microscale thermophoresis(MST)and isothermal titration calorimetry(ITC)were used to study in vitro interaction between CMV 2b protein and antiviral small molecule compounds to screen highly active antiviral small molecule compounds.On the other hand,agrobacterium-mediated method,real-time quantitative polymerase chain reaction(q PCR)and western blotting(WB)were used to verify its activity in vivo,and site-directed mutagenesis was applied in the preliminary verification of the key binding sites between CMV 2b protein and highly active antiviral small molecule compounds.The experimental results showed that the fusion expression protein of CMV 2b was successfully obtained,and the compound LP11 that has good binding with CMV2 b protein was screened by MST combined with ITC experiment.q PCR and WB results confirmed that LP11 can effectively inhibit CMV infection in tobacco,at the transcription and translation levels.The mutation results shows that the 15 th leucine and 18 th methionine at the N-terminal of CMV 2b protein are the key binding sites of compound LP11,which provided a theoretical basis for screening and developing anti-CMV agents.