Construction And Reactionogenicity Analysis Of Recombinant Fowlpox Viruses Expressing VP3 Gene Of Goose Parvovirus

Goose Parvovirus (Goose Parvovirusis, GP) was gave rise to goose parvovirus (Goose Parvovirus, GPV) goslings caused acute, subacute and contacted frequently infectious diseases. GPV genome contained three structural proteins (VP1, VP2 and VP3). However, the VP3 protein was main component of the viral capsid, about 80% of the total protein, which could stimulate the organism produced neutralized antibodies, so GPV VP3 was the first choice genes developed gene live vector vaccine. And it played an important role in respects of transformated fowl pox virus, gene expression, live vector vaccine development as well as disease treatment. Yet it was deleted the genes caused avian pox virus, but retained the replication activity with high security, sustained expression exogenous target gene, which produced immunity for a long time after once vaccination. As made it a live vector vaccine most widely used expression vector.Today, it generally take traditional ways to GP Prevented Vaccine adult goose lead goslings get immune protection from maternal antibodies in native and abroad,it also had a direct immune goslings get immune protection. However, attenuat ed vaccine strain, inactivated vaccines and other traditional vaccines had deficiency such as reversion of virulence, inactivation was not complete, epidemic diffusion, undistinguished between naturally infected with immune deficiencies, the u rgent need to develop a safe effective new vaccine. Up to now, the research of GPV protective antigen gene of avian pox virus live vector vaccine has been preliminary stage, there was a big space research in fowl pox vaccines recombination viral vectors in GPV.In this study, virulent strain (YBLJ strain) GPV-VP3 gene isolated from Yanbian was a protective antigen gene to construct VP3 gene to the downstream of pSY538 vector by BamH I site, construct pSY538-VP3 vector; through one step cloning method connect Lac Z gene which was smoothing terminal to the Sma I site of pSY538-VP3, construct pSY538-VP3-Lac Z vector; use Not I to cut the expression cassette of VP3 and Lac Z, inserted into the pSY681 vector, the recombination express GPV-VP3 gene of avian poxvirus transfered vector pSY681-GPV-VP3. Identification the colligated direction by sequence and connect by restriction enzyme, transfected prior infection S-FPV-017 parent FPV chicken embryo fibroblasts (CEF), by homologous arms crossing-over. Screening and purification by plaque assay, get purification recombinant fowlpox virus through 10 times screening, ultimately IFA and western-blot identification expressed recombination fowl pox virus GPV-VP3 gene expressed production and bad preliminary reactionogenicity analysis.This study further GPV-VP3 recombination Experimental study of immune fowl pox virus vector vaccine, as well as explored the GPV-VP3 recombination immune protection and immuned protection mechanisms fowl pox virus vector vaccine foundation.