Expression,Purification and Characterization of A33R,L1R,B5R and A27L Proteins of Monkeypox Virus
This paper intends to explore the prokaryotic expression of GST-agged fusion proteins A33R,L1R,B5R and A27L,and conduct a brief analysis of some biological characteristics of these four proteins,laying the foundation for monkeypox virus antigen preparation and vaccine preparation.Recombinant fu-sion protein expression plasmid was constructed with PGEX-6P-1 as vector.The recombinant plasmid was transformed into E.coli DE3 competent cells and induced to express and purify in vitro.The purified pro-tein was subjected to SDS-PAGE electrophoresis,Coomassie brilliant blue staining analysis and WB verifi-cation.Finally,the biological characteristics of the four proteins were analyzed by bioinformatics methods,and the physical and chemical properties of the four proteins were analyzed by ProtParam tool in ExPAsy online software.The ProtScale tool in ExPAsy online software was used to analyze the hydrophilicity and hydrophobicity of the four proteins.The signal peptides of the four proteins were analyzed by SignalP4.1 service online software.Four recombinant expression plasmids were successfully constructed.Coomassie brilliant blue and WB verification showed that the target protein with GST tag was successfully induced and purified.Bioinformatics analysis showed that four proteins were hydrophobic proteins,except B5R,A33R,L1R and A27L were secretory proteins.In this study,four proteins of monkeypox virus were puri-fied by prokaryotic expression,and the biological characteristics of these four proteins were analyzed by bioinformatics,which provided a theoretical basis for antigen preparation and vaccine preparation of mon-keypox virus.
万方数据
维普
