Rescue and identification of recombinant fowl adenovirus 4 expressingσB and σC of genotype-Ⅰ subcluster avian reovirus
Inorder to obtain a recombinant fowl adenovirus 4(FAdV-4)strain that could express the σB and σC of avian reovirus Ⅰ-sub(ARV GCI-sub),a novel Fosmid clay rescue system was used to rescue the virus using the fowl adenovirus rHN20 as the vector and the recombinant virus rHN20-r1966-ARV GCI-sub ex-pressing σB and σC proteins was constructed.The successful rescue of recombinant virus was proved by PCR,indirect immunofluorescence assay(IFA),Wes tern-blot,and the kinetics of replication in vi tro.The results showed that σB and σC gene of ARV GCI sub were detected in the LMH cells infected with recombi-nant virus by the PCR,Fiber protein of FAdV-4,σB and σC protein of ARV GCI sub were detected in the LMH cells infected with recombinant virus by the Western-blot experiment,and σB and σC protein of ARV GCI sub were specifically detected in the LMH cells infected with recombinant virus by the IFA experiment.The recombinant virus showed no significant difference in replication ability between the recombinant virus and the parent virus,indicating successful rescue of recombinant virus rHN20-r1966-ARV GCI-sub.The construction of recombinant virus rHN20-r1966-ARV GCI-sub provides technical support for ARV pre-vention and control vaccines targeting σB and σC protein of ARV generation at the same time,and lays a foundation for the combined prevention and control of ARV and FAdV-4.
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