Expression of nonstructural proteins in Japanese encephalitis virus and their interaction with host hnRNP K
[Objective]This study aims to discuss the interaction between JEV non-structural protein NS1,NS2a,NS3,NS5 and host hnRNP K protein,so as to study the relationship between JEV virus replication and host protein.[Methods]First,non-structural protein granules were constructed,and then the constructed non-structural protein plasmids were transfected into Vero cells,the protein expression was observed under fluorescence microscope for 24 h,and then simultaneously transfected into Vero cells with pCMV-MYC-hnRNP K plasmid,which was cultured for 24 h,and cell protein samples were collected.The relevant steps of Co-IP experiment were carried out,and the results were presented by Western-blot experiment to obtain the test results.[Results]The findings demonstrated successful expression of the unstructured protein plasmid in Vero cells as per this study.Co-transfection with the hnRNP K protein-expressing plasmid,followed by co-immunoprecipitation(Co-IP)test and Western blot analysis,revealed an interaction between JEV NS1 and hnRNP K.However,no interaction was observed be-tween JEV NS2a,NS3,NS5 and host hnRNP K.[Conclusion]The interaction between JEV NS1 protein and host protein hnRNP K further broadens the thinking of the study of JEV nonstructural proteins,provides a foundation for the study of the life cycle of JEV virus in host cells,and provides a new direction for the further study of JEV replication.
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