Co-expression Of Goose Parvovirus VP3 Gene And Goose Paramyxovirus F Gene In Recombinant Avipox Virus

Goose parvovirus and goose paramyxovirus are the pathogens of Gosling plaque and Geese'paramyxovirus disease, both of them are violent fatal infectious diseases of goose existing nearlyaround the world. The diseases are contagious among goose of ages and breeds. High death rate ofinfected gooses, especially goslings, is the feature of these two diseases. The genome of gooseparvovirus encode three proteins which are VP1, VP2 and VP3 respectively. VP3 is the mainstructural protein with highly conservatism,which can induce neutralization antibodies.So VP3 is themain immunogenicity protein of goose parvovirus. Goose paramyxovirus is the member ofgroup I paramyxovirus, has the typical character of geneâ…¦type Newcastal diseasevirus(NDV). The peplos of goose paramyxovirus is composed by two proteins.One is fusionprotein(F protein) which mediates the fusion of virus peplos and cytomembrane. F protein plays avary important role in the immunoprotection of this disease.The control of GPV and GPMV mainly depends on traditional vaccines currently in ourc0untry.But the traditional vaccines have some shortcomings in practical application,as the highcost, easily spreading viruses, interfering monitoring and so on. With the development of geneticengineering technology towards the research of vaccine, genetic engeneering vaccine are becomingmore and more popular, which is supposed to be more effective and safer as vaccines, and therecombinant fowlpox virus is a tapical example.The Lipofectamine (TM)~2000-PLUS~+ Reagent was applied for transferring the vectorpSY681-VP3(GPV)-F(GPMV)-LacZ constructed previously to the CEF cell infected with avipoxvirus(FPV-017). Recombinant avipox viruses were selected by blue plaque screening based on theexpressing of reporter LacZ gene, purified by five cycles cloning of blue plaque. The results ofidentification by PCR indicated that VP3 gene and F gene have been inserted into the genome ofavipox virus; and expression of VP3 gene and F gene was detected by Dot-ELISA andindirect-immunofluorescence test, the results indicated that recombinant avipox virus could expressthe two foreign genes efficiently ,and the two proteins possessed reactionogenicity. This studyprovided the theoretical and practical foundation for recombinant avipox virus genetic engineeringvaccine co-expressing two foreign genes, as well as provided substance preparation for preventionthe GPV and GPMV.