减蛋综合征病毒纤维蛋白和六邻体蛋白可溶性表达条件的优化OA

To achieve soluble prokaryotic expression of Fiber,Penton and Hexon proteins of Egg Drop Syn-drome Virus(EDSV)strain AV127,the genes encoding these proteins were cloned into the multiple clo-ning site of the prokaryotic expression vector pET-32a(+),generating recombinant plasmids pET-32a(+)-Fiber,pET-32a(+)-Hexon,and pET-32a(+)-Penton.Then,these three recombinant plasmids were transformed into Rosetta(DE3)for induction expression,and the soluble expression conditions of Fiber and Hexon proteins were further optimized to achieve the maximum soluble expression level.The SDS-PAGE i-dentification results showed that Penton protein had the highest soluble expression level,accounting for 95.30％of the total bacterial protein.Hexon protein increased its soluble expression level from 63.29％to 83.73％by optimizing the concentration of IPTG.For Fiber protein,by optimizing the expression vector,in-duction temperature,time,and expression strain,the soluble expression level was increased from 28.54％to 75.04％,achieving the maximum soluble expression of Fiber,Penton,and Hexon proteins.This study suc-cessfully established an efficient expression system for the major structural protein of EDSV,laid a solid foundation for subsequent protein purification and functional studies,and also had important reference val-ue for the prokaryotic expression of other viral structural proteins.