Eukaryotic-Expressed Chicken CCCH-Type Zinc Finger Antiviral Protein Resist Infection Of Four Virus

Natural antiviral factor is a host endogenous immune factor with antiviral effect,which is an important way to resist virus invasion formed in the repeated struggle between host and virus,and plays an important role in the immune system of the body studies have shown that zinc-finger protein is also a natural host endogenous immune factor that regulates cytokine expression such as interleukin,interferon,and directly regulates lymphocyte growth and differentiation.chZAP is a host anti-mouse leukemia virus(MLV)protein factor screened by a functional genomic method cloned in 2002 Overexpression of chZAP in murine fibroblasts can inhibit MLV replication.Studies have shown that zinc finger protein is also an innate antiviral factor and plays an important role in the body's immune system.Zinc-finger protein can regulate the expression of cytokines such as interleukin and interferon,and directly regulate the growth and differentiation of lymphocytes.During the innate immune response,zinc-finger proteins play an antiviral effect directly or indirectly.In recent years,studies have found that chZAP has a significant inhibitory effect on Ross River virus,Venezuelan equine encephalitis virus,and Ebola virus,but it has no significant inhibitory effect on herpes virus,type 1 vesicular stomatitis virus,and yellow fever virus.It shows that chZAP does not have broad-spectrum antiviral properties Nonetheless,little is known about its antiviral activity against avian viruses,and its structure and function remain to be studied.It has been proved that chZAP interacts with viruses by degrading viral mRNA and has antiviral effects,and participates in T cell immune response.If we can study the function of avian zinc-finger protein,screen its target virus,and determine its antiviral spectrum,it may provide new ideas for the development of new natural immune drugs or prevention and control of avian diseaseThe chZAP gene was amplified,the target gene was sublinked to the eukaryotic expression vector pBacPAK8,the recombinant baculovirus plasmid was constructed,the recombinant baculovirus plasmid was transfected into the sf9 cells,the P1 generation baculovirus was connected to the sf9 cells,the generation baculovirus was prepared,the cell lysate and the cell culture supernatant were collected for eukaryotic expression of the target protein.the results showed that the target protein was expressed in both cell lysate and cell culture supernatant.chZAP protein was obtained by purification of the expressed recombinant protein by nickel column method.The two proteins were first validated in vitro The specific methods were as follows:culture DF-1 cells,inoculation NDV?MDV?IBDV?REV,determination of cell infection virus,access to the appropriate concentration of chZAP protein,after a period of culture,qPCR and western blot to detect the changes of virus load and protein expression in different test groups.The results showed that the chZAP protein had no obvious inhibitory effect on MDV,but on NDV?IBDV?REV to a certain extentThe purpose of this study was to analyze the effect of over-expressed chZAP on virus replication,with the aim of identifying the target virus type for which chZAP exerts antiviral effects,and clarifying the antiviral spectrum of chZAP.In theory,by analyzing the target virus types of chZAP,it can provide a reference for revealing the anti-virus mechanism of chZAP.In practice,it guides the development of new antiviral drugs to a certain extent.